Methods and compositions for producing plants and microorganisms that express feedback insensitive threonine dehydratase/deaminase

ABSTRACT

The present invention relates to methods and materials in the field of molecular biology and the regulation of polypeptide synthesis through genetic engineering of plants and/or microorganisms. More particularly, the invention relates to newly-isolated nucleotide sequences, nucleotide sequences having substantial identity thereto and equivalents thereof, as well as polypeptides encoded thereby. The invention also involves the introduction of foreign nucleotide sequences into the genome of a plant and/or microorganism, wherein the introduction of the nucleotide sequence effects an increase in the transformant&#39;s resistance to toxic isoleucine structural analogs. Inventive sequences may therefore be used as excellent molecular markers for screening successful transformants, thereby replacing antibiotic resistance genes used in the prior art. Transformants harboring a nucleotide sequence comprising a promoter operably linked to an inventive nucleotide sequence demonstrate increased levels of isoleucine production, thereby providing an improved nutrient source.

REFERENCES TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 60/052,096, filed Jul. 10, 1997 and entitled cDNA CLONE SEQUENCE OF THREONINE DEHYDRATASE/DEAMINASE FROM ARABIDOPSIS THALIANA; and U.S. Provisional Application No. 60/074,875, filed Feb. 17, 1998 and entitled THE MOLECULAR BASIS OF L-O-METHYLTHREONINE RESISTANCE ENCODED BY THE omr1 ALLELE OF LINE GM11b OF ARABIDOPSIS THALIANA; both of which are hereby incorporated by reference herein in their entirety.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to methods and materials in the field of molecular biology and to the utilization of isolated nucleotide sequences to genetically engineer plants, and/or microorganisms. More particularly, the invention relates in certain preferred aspects to novel nucleotide sequences and uses thereof, including their use in DNA constructs for transforming plants, fungi, yeast & bacteria. The nucleotide sequences are particularly useful as selectable markers for screening plants and/or microorganisms for successful transformants and also for improving the nutritional value of plants.

[0004] 2. Introduction and Discussion of Related Art

[0005] Threonine dehydratase/deaminase (“TD”) is the first enzyme in the biosynthetic pathway of isoleucine, and catalyzes the formation of 2-oxobutyrate from threonine (“Thr”) in a two-step reaction. The first step is a dehydration of Thr, followed by rehydration and liberation of ammonia. All reactions downstream from TD are catalyzed by enzymes that are shared by the two main branches of the biosynthetic pathway that lead to the production of the branched-chain amino acids, isoleucine (“Ile”), leucine (“Leu”), and valine (“Val”). An illustration of the biosynthetic pathway is set forth in FIG. 1. The cellular levels of Ile are controlled by negative feedback inhibition. When the cellular levels of Ile are high, Ile binds to TD at a regulatory site (allosteric site) that is different from the substrate binding site (catalytic site) of the enzyme. The formation of this Ile-TD complex causes conformational changes to TD, which prevent the binding of substrate, thus inhibiting the Ile biosynthetic pathway.

[0006] It is known that certain structural analogs of Ile exist which are toxic to a wide variety of plants and microorganisms. It is believed that these Ile analogs are toxic because cells incorporate the analogs into polypeptides in place of Ile, thereby synthesizing defective polypeptides. In this regard, L-O-methylthreonine (“OMT”) was reported in 1955 to be a structural analog of Ile that inhibits growth of manmalian cell cultures by inhibiting incorporation of Ile into proteins. (Rabinovitz M, et al., Steric relationship between threonine and isoleucine as indicated by an antimetabolite study. J Am Chem Soc 77:3109-3111 (1955).) It is believed that the same phenomenon explains growth inhibition, which is caused by other structural analogs of Ile such as, for example, thiaIle.

[0007] Certain strains of bacteria and yeast and certain plant lines have been identified which are resistant to the toxicity of the above-noted Ile structural analogs, and this resistance has been attributed to a mutation in the TD enzyme. The mutated TD apparently features a loss or decrease of Ile feedback sensitivity (referred to herein as “insensitivity”). As a result of this insensitivity, cells harboring insensitive TD produce increased amounts of Ile, thereby outcompeting the toxic Ile analog during incorporation into cellular proteins. For example, resistance to thiaIle has been associated in certain strains of bacteria and yeast with a loss of feedback sensitivity of TD to Ile. In Rosa cells, resistance to OMT was also associated with a TD that had reduced sensitivity to feedback inhibition by Ile. Being in tissue culture and having high ploidy level, however, it was not possible to determine the genetic basis of feedback insensitivity to Ile in the Rosa variant, the only known plant mutated with an Ile-insensitive TD.

[0008] Turning to a field of research where the present invention finds advantageous application, selectable markers are widely used in methods for genetically transforming cells, tissues and organisms. Such markers are used to screen cells, most commonly bacteria, to determine whether a transformation procedure has been successful. As a specific example, it is widely known that constructs for transforming a cell may include as a selectable marker a nucleotide sequence that confers antibiotic resistance to the transformed cell. As used herein in connection with cells and plants, the terms “transformed” and “transgenic” are used interchangeably to refer to a cell or plant expressing a foreign nucleotide sequence introduced through transformation efforts. The term “foreign nucleotide sequence” is intended to indicate a sequence encoding a polypeptide whose exact amino acid sequence is not normally found in the host cell, but is introduced therein through transformation techniques. After transformation, the cells may be contacted with an antibiotic in a screening procedure. Only successful transformants, i.e., those which possess the antibiotic resistance gene, survive and continue to grow and proliferate in the presence of the antibiotic. This techniques provides a manner whereby successful transformants may be identified and propagated, thereby eliminating the time consuming and costly alternative of growing and working with cells which were not successfully transformed.

[0009] The above-described screening technique is becoming less advantageous, however, because, due to prolonged exposure to antibiotics, an ever-increasing number of naturally-occurring microorganisms are developing antibiotic resistance by spontaneous mutation. The reliability of this screening technique is therefore compromised because the continuous exposure to antibiotics causes microorganisms that are not transformed to develop spontaneo'us mutations that confer antibiotic resistance.

[0010] In addition to the decreasing viability of this screening technique, the overuse of antibiotics, and the resulting resistance spontaneously developed by microorganisms, is a growing medical concern as the efficacy of antibiotics in fighting bacterial infections is decreasing. Many infections—including meningitis—no longer respond well to drugs that once worked well against them. This phenomenon is attributed largely to the overuse of antibiotics, both as drugs and as a laboratory screening tool, and the resulting antibiotic resistance of a growing number of microorganisms. As an example, the bacteria that causes meningitis once was routinely controlled with ampicillin, a commonly prescribed antibiotic and an antibiotic very heavily used in screening transformed bacterial cells for resistance as a selectable marker. Now, however, about 20 percent of such infections are resistant to ampicillin.

[0011] The present invention addresses the aforementioned problems in screening genetic transformants and provides nucleotide sequences which may be advantageously used as selectable markers, and which may be inserted into the genome of a plant or microorganism to provide a transformed plant or microorganism. Such a transformed plant or microorganism advantageously exhibits significantly increased levels of Ile synthesis and synthesis of intermediates of the Ile biosynthetic pathway and is therefore also capable of surviving in the presence of a toxic Ile analog.

SUMMARY OF THE INVENTION

[0012] The present invention provides nucleotide sequences, originally isolated and cloned from Arabidopsis thaliana, which encode feedback insensitive TD that may advantageously be used to transform a wide variety of plants, fungi, bacteria and yeast. Inventive forms of TD are not only insensitive to feedback inhibition by isoleucine, but are also insensitive to structural analogs of isoleucine that are toxic to plants and microorganisms which synthesize only wild-type TD. Therefore, inventive nucleotide sequences encoding mutated forms of TD can be used to create cells that are insensitive to compounds normally toxic to cells expressing only wild-type TD enzymes. In this regard, an inventive nucleotide sequence may be used in a DNA construct to provide a biochemical selectable marker

[0013] One aspect of the present invention is identification, isolation and purification of a gene encoding a wild-type form of TD. The DNA sequence thereof can be used as disclosed herein to determine the complete amino acid sequence for the protein encoded thereby and thus allow identification of domains found therein that can be mutated to produce additional TD proteins having altered enzymatic characteristics. In another aspect of the invention, there are provided isolated and purified polynucleotides, the polynucleotides encoding a mutated form of TD, or a portion thereof, as disclosed herein. For example, the invention provides isolated polynucleotides comprising the sequence set forth in SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4, nucleotide sequences having substantial identity thereto, and nucleotide sequences encoding TD variants of the invention. Also provided are isolated polypeptides comprising the amino acid sequence set forth in SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4 and variants thereof selected in accordance with the invention.

[0014] In an alternate aspect of the invention, there is provided a chimeric DNA construct comprising a promoter operably linked to a nucleotide sequence encoding a threonine dehydratase/deaminase that is substantially resistant to feedback inhibition. In a cell harboring the construct, the nucleotide sequence can be transcribed to produce mRNA and said mRNA can be translated to produce either mature, mutated TD or a precursor mutated TD protein, said protein being functional in said cell. Also provided, therefore, is a vector useful for transforming a cell, and plants and microorganisms transformed therewith, the vector comprising a DNA construct selected in accordance with the invention. In alternate aspects of the invention, there are provided cells and plants having incorporated into their genome a foreign nucleotide sequence operably linked to a promoter, the foreign sequence comprising a nucleotide sequence having substantial identity to a sequence set forth herein or a foreign nucleotide sequence encoding an inventive polypeptide.

[0015] In another aspect of the invention, there is provided a method comprising incorporating into a plant's genome an inventive DNA construct to provide a transformed plant; wherein the transformed plant is capable of expressing the nucleotide sequence.

[0016] Yet another aspect of the invention is the production and propagation of cells transformed in accordance with the invention, wherein the cells express a mutated TD enzyme, thus making the cells resistant to feedback inhibition by isoleucine, and resistant to molecules that are toxic to a cell producing only the wild-type TD enzyme. In this regard, there is provided a method comprising providing a vector featuring a promoter operably linked to a nucleotide sequence encoding a threonine dehydratase/deaminase that is resistant to feedback inhibition, wherein the promoter regulates expression of the nucleotide sequence in a host plant cell; and transforming a target plant with the vector to provide a transformed plant, the transformed plant being capable of expressing the nucleotide sequence. Plants transformed in accordance with the invention have within their chloroplasts a mature, mutated form of TD, which renders the cells resistant to toxic Ile analogs. Also provided are transformed plants obtained according to inventive methods and progeny thereof.

[0017] Also provided is a method for screening potential transformants, comprising (1) providing a plurality of cells, wherein at least one of the cells has in its genome an expressible foreign nucleotide sequence selected in accordance with the invention; and (2) contacting the plurality of cells with a substrate comprising a toxic isoleucine structural analog; wherein cells comprising the, expressible foreign nucleotide sequence are capable of growing in the substrate, and wherein cells not comprising the expressible foreign nucleotide sequence are incapable of growing in the substrate.

[0018] In another aspect of the invention, there is provided a construct comprising a primary nucleotide sequence to be introduced into the genome of a target cell, tissue and/or organism, and further comprising a biochemical selectable marker selected in accordance with the invention. This aspect of the invention may be advantageously used to transform a wide variety of cells, including microorganisms and plant cells. After introducing the DNA construct, which also includes an appropriate promoter and such other regulatory sequences as may be selected by a skilled artisan, into a target plant or microorganism, the plant or microorganism may be grown in a substrate comprising a toxic isoleucine analog (a “toxic substrate”), thereby providing a mechanism for the early determination whether the transformation was successful. Where a plurality of plants or microorganisms are transformed, placing potential transformants into a toxic substrate provides an early screening step whereby successful transformants may be identified. It is readily understood by a person skilled in the relevant field, in view of the present specification, that successful transformants will grow normally in the toxic substrate by virtue of expression of the insensitive TD; however, unsuccessfully transformed plants and/or microorganisms will die due to the toxic effect of the substrate. Transformed plants may thereby be identified quickly in accordance with the invention, and transformed microorganisms may be identified in accordance with the invention without using antibiotic resistance genes.

[0019] In another aspect of the invention, there is provided a method for reliably incorporating a first, expressible, foreign nucleotide sequence into a target cell, comprising providing a vector comprising a promoter operably linked to a first primary nucleotide sequence and a second nucleotide sequence selected in accordance with the invention, the second sequence encoding an insensitive TD enzyme; transforming the target cell with the vector to provide a transformed cell; and contacting the cell with a substrate comprising L-O-methylthreonine; wherein successfully transformed cells are capable of growing in the substrate, and wherein unsuccessfully transformed cells are incapable of growing in the substrate.

[0020] In an alternate aspect of the invention, there is provided a method for growing a plurality of plants in the absence of undesirable plants, such as, for example, weeds, the method comprising providing a plurality of plants, each having in its genome a foreign nucleotide sequence comprising a promoter operably linked to a nucleotide sequence selected in accordance with the invention; growing the plurality of plants in a substrate; and introducing a preselected amount of an isoleucine structural analog into the substrate.

[0021] TD enzymes described herein function in the chloroplasts of a plant cell. Therefore, it is readily appreciated by a skilled artisan that a nucleotide sequence inserted into a plant cell will necessarily encode a precursor TD peptide. Thus, chimeric DNA constructs are described herein that comprise a first nucleotide sequence encoding a mature mutated form of TD and a second nucleotide sequence encoding a chloroplast transit peptide of choice, the second sequence being functionally attached to the 5′ end of the first sequence. Expression of the chimeric DNA construct results in the production of a mutated precursor TD enzyme that can be translocated to a chloroplast. The presence of a mature mutated TD in the chloroplast results in a plant cell having characteristics described herein.

[0022] It is an object of the present invention to provide isolated nucleotide sequences, which may be introduced into the genome of a plant or microorganism to increase the ability of the plant or microorganism to synthesize Ile and intermediates of the Ile biosynthetic pathway.

[0023] Additionally, it is an object of the invention to provide nucleotide sequences, which may be used as excellent biochemical selectable markers for identifying successful transformants in genetic engineering protocols.

[0024] It is also an object of the invention to provide a novel, efficient, selective, environmentally-friendly herbicide system.

[0025] Further objects, advantages and features of the present invention will be apparent from the detailed description herein.

BRIEF DESCRIPTION OF THE FIGURES

[0026] Although the characteristic features of this invention will be particularly pointed out in the claims, the invention itself, and the manner in which it may be made and used, may be better understood by referring to the following description taken in connection with the accompanying figures forming a part hereof.

[0027]FIG. 1 illustrates the biosynthetic pathway of the branched-chain amino acids valine, leucine and isoleucine.

[0028]FIG. 2 sets forth the alignment of the amino acid sequence of TD of tomato and chickpea. C regions are highly conserved regions of the catalytic site of TD while R regions are highly conserved regions of the regulatory site of TD. Also shown are the locations of the degenerate oligonucleotide primers TD205 and TD206 used to PCR-amplify an Arabidopsis TD genomic DNA fragment

[0029]FIG. 3 sets forth the structure and degree of degeneracy of the two oligonucleotide primers TD205 and TD206 used in amplifying an Arabidopsis genomic DNA fragment of the TD gene omr1. TD205 is anchored with an Eco RI site (underlined) at its 5′ end and TD206 is anchored with a Hind III site (underlined) at its 5′ end.

[0030]FIG. 4 sets forth the DNA sequence of clone 23 (pGM-td23) isolated from a cDNA library of the mutated line GM11b (omr1/omr1) of Arabidopsis thaliana.

[0031]FIG. 5 sets forth the nucleotide sequence and the predicted amino acid sequence of clone 23 as isorated from the cDNA library constructed from line GM11b of Arabidopsis (omr1/omr1). The TD insert in clone 23 is in pBluescript vector between the Eco RI and Xho I sites. An open reading frame (top reading frame) was observed which showed an ATG codon at nucleotide 166 and a termination codon at nucleotide 1801.

[0032]FIG. 6a depicts the structure of the expression vector pCM35S-omr1 used in the transformation of wild-type Arabidopsis thaliana and which expressed a mutated form of TD capable of conferring resistance to the toxic analog L-O-methylthreonine upon transformants.

[0033]FIG. 6b sets forth the nucleotide sequence and the predicted amino acid sequence of the chimeric mutant omr1 expressing resistance to L-O-methylthreonine in transgenic Arabidopsis plants that have been transformed with the expression vector pCM35s-omr1 (shown in FIG. 6a). The total length of the fusion (chimeric) mutant TD expressed in transgenic plants was 609 amino acid residues. The first 9 amino-terminal residues start by methionine encoded by a start codon (ATG) furnished by the 3′ end of the nucleotide sequence of CaMV 35s promoter linked to the omr1 insert of clone 23. The following 15 amino acid residues are generated by the nucleotide sequence of the polylinker region from the multiple cloning site of the vector and finally the remaining 585 amino acid residues are encoded by the omr1 mutant allele of Arabidopsis as present in clone 23. The first residue of the 585 amino acid long portion encoded by omr1 in pCM35s-omr1 corresponds to threonine (Thr) which is the amino-terminal residue number 8 of the full length omr1 cDNA shown in FIGS. 8 and 9 and SEQ ID NO:2.

[0034]FIG. 7 is the nucleotide sequence of the full length cDNA of the omr1 allele encoding mutated TD. The total length of the cDNA of omr1 is 1779 nucleotides including the stop codon.

[0035]FIG. 8 is the predicted amino acid sequence of the mutated TD encoded by omr1. The total length of the TD protein encoded by omr1 is 592 amino acids.

[0036]FIG. 9 is the nucleotide sequence and the predicted amino acid sequence encoded by the mutated allele omr1 of line GM11b of Arabidopsis thaliana.

[0037]FIG. 10 is the nucleotide sequence of the full length cDNA of the wild type allele OMR1 encoding wild type TD.

[0038]FIG. 11 is the predicted amino acid sequence of the wild type TD encoded by OMR1.

[0039]FIG. 12 is the nucleotide sequence and the predicted amino acid sequence encoded by the wild type allele OMR1 of Arabidopsis thaliana Columbia wild type.

[0040]FIG. 13 sets forth the multi-alignment of the deduced amino acid sequence of the wild-type TD of Arabidopsis thaliana reported in this disclosure with that from other organisms obtained from GenBank with the following accession numbers: 940472 for chickpea; 10257 for tomato; 401179 for potato; 730940 for yeast 1; 134962 for yeast 2; 68318 for E. coli biosynthetic; 135723 for E. coli catabolic; 1174668 for Salmonella typhimurium. The megalign program of the Lasergene software, DNASTAR Inc., Madison, Wis. was used.

[0041]FIG. 14 is a portion of the DNA sequencing gel comparing the nucleotide sequence of the mutated omr1 allele and its wild-type allele OMR1 and showing the base substitution C (in OMR1) to T (in omr1) at nucleotide residue 1495 starting from the beginning of the coding sequence. The arrow is pointing to the base substitution.

[0042]FIG. 15 depicts the point mutation in omr1 at nucleotide residue 1495, predicting an amino acid substitution, from arginine (R) to cysteine (C) at amino acid residue 499 at the TD level.

[0043]FIG. 16 sets forth the amino acid sequence at the regulatory region R4 of TD encoded by mutated omr1 and wild type OMR1) alleles of Arabidopsis thaliana compared to that from several organisms. The arrow points to the mutated amino acid residue in omr1.

[0044]FIG. 17 is a portion of the DNA sequencing gel comparing the nucleotide sequence of the mutated omr1 allele and its wild-type allele OMR1 and showing the base substitution G (in OMR1) to A (in omr1) at nucleotide residue 1631. The arrow is pointing to the base substitution.

[0045]FIG. 18 depicts the point mutation in omr1 at nucleotide residue 1631, predicting an amino acid substitution, arginine (R) to histidine (H) at amino acid residue 544 at the TD level.

[0046]FIG. 19 sets forth the amino acid sequence at the regulatory region R6 of TD encoded by mutated omr1 and wild type OMR1 alleles of Arabidopsis thaliana compared to that from several organisms. The arrow points to the mutated amino acid residue in omr1.

DETAILED DESCRIPTION OF THE INVENTION

[0047] For purposes of promoting an understanding of the principles of the invention, reference will now be made to particular embodiments of the invention and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the invention, and such further applications of the principles of the invention as described herein being contemplated as would normally occur to one skilled in the art to which the invention pertains.

[0048] As disclosed above, the present invention relates to methods and compositions for obtaining transformed cells, said cells expressing therein a mutated form of threonine dehydratase/deaminase (“TD”). More particularly, the invention provides isolated nucleotide sequences encoding mutated TD-functional polypeptides (“mutated TD”) which are resistant to Ile feedback inhibition and are resistant to the toxic effect of Ile analogs. These inventive nucleotide sequences can be incorporated into vectors, which in turn can be used to transform cells. Such transformation can be used, for instance, for purposes of providing a selectable marker, to increase plant nutritional value or to increase the production of commercially-important intermediates of the isoleucine biosynthetic pathway. Expression of the mutated TD results in the cell having altered susceptibility to certain enzyme inhibitors relative to cells having wild-type TD only. These and other features of the invention are described in further detail below.

[0049] One feature of the present invention involves the discovery, isolation and characterization of a gene sequence from Arabidopsis thaliana, designated omr1, which encodes a surprisingly advantageous mutated form of the enzyme TD. Aspects of the present invention thus relate to nucleotide sequences encoding mutated forms of TD, which sequences may be introduced into target plant cells or microorganisms to provide a transformed plant or microorganism having a number of desirable features. The mutated forms of TD, unlike wild-type TD, are resistant to negative feedback inhibition by isoleucine (“Ile”) and transformed cells are resistant to molecules which are toxic to cells that do not express feedback insensitive TD. Therefore, transformants harboring an expressible inventive nucleotide sequence demonstrate increased levels of isoleucene production and increased levels of production of intermediates in the Ile biosynthetic pathway, and the transformants are resistant to Ile structural analogs which are lethal to non-transformants, which express only wild-type TD.

[0050] The present invention relates in another aspect to amino acid sequences that comprise functional, feedback-insensitive TD enzymes. The term “amino acid sequence” is used herein to designate a plurality of amino acids linked in a serial array. Skilled artisans will recognize that through the process of mutation and/or evolution, polypeptides of different lengths and having differing constituents, e.g., with amino acid insertions, substitutions, deletions, and the like, may arise that are related to a sequence set forth herein by virtue of amino acid sequence homology and advantageous functionality as described in detail herein. The term “TD enzyme” is used to refer generally to a wild-type TD amino acid sequence, to a mutated TD selected in accordance with the invention, and to variants of each which catalyzes the reaction of threonine to 2-oxobutyrate in the Ile biosynthetic pathway, as described herein. For purposes of clarity, the wild-type form is distinguished from a mutated form, where necessary, by usage of the terms “wild-type TD” and “mutated TD.”

[0051] It is not intended that the present invention be limited to the specific sequences set forth herein. It is well known that plants and microorganisms of a wide variety of species commonly express and utilize analogous enzymes and/or polypeptides which have varying degrees of degeneracy, and yet which effectively provide the same or a similar function. For example, an amino acid sequence isolated from one species may differ to a certain degree from the wild-type sequence set forth in SEQ ID NO:1, and yet have similar functionality with respect to catalitic and regulatory function. Amino acid sequences comprising such variations are included within the scope of the present invention and are considered substantially similar to a reference amino acid sequence. It is believed that the identity between amino acid sequences that is necessary to maintain proper functionality is related to maintenance of the tertiary structure of the polypeptide such that specific interactive sequences will be properly located and will have the desired activity. While it is not intended that the present invention be limited by any theory by which it achieves its advantageous result, it is contemplated that a polypeptide including these interactive sequences in proper spatial context will have good activity, even where alterations exist in other portions thereof.

[0052] In this regard, a TD variant is expected to be functionally similar to the wild-type TD set forth in SEQ ID NO:1, for example, if it includes amino acids which are conserved among a variety of species or if it includes non-conserved amino acids which exist at a given location in another species that expresses functional TD. FIG. 13 sets forth an amino acid alignment of TD polypeptides of a number of species. Two significant observations which may be made based upon FIG. 13 are (1) that there is a high degree of conservation of amino acids at many locations among the species shown, and (2) a number of insertions, substitutions and/or deletions are represented in the TD of certain species and/or strains, which do not eliminate the dual functionality of the respective TD enzymes. For example, on Page 4 of FIG. 13, Regulatory Region 4 (“R4”) of wild-type Arabidopsis is depicted which comprises the following sequence (corresponding to the underlying three-letter codes numbered as set forth in SEQ ID NO:1):  V   N   L   T   T   S   D   L   V   K   D   H   L   R   Y   L   M   G   G Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp His Leu Arg Tyr Leu Met Gly Gly 486             490                 495                 500

[0053] The degeneracy shown in FIG. 13 in this portion of the sequence provides examples of substitutions which may be made without substantially altering the functionality of the wild-type sequence set forth in SEQ ID NO:1. For example, it is expected that the Asp (“D”) at position 492 could be substituted with a Glu (“E”) and that the Leu (“L”) at position 493 could be substituted with a Met (“M”) without substantially altering the functionality of the amino acid sequence.

[0054] The following sets forth a plurality of sequences of R4, depicted such that acceptable substitutions are set forth at various amino acid locations. The sequences encompassed thereby are expected to exhibit similar functionality to the corresponding portion of SEQ ID NO:1. A slash (“/”) between two or in a series of amino acids indicates that any one of the amino acids indicated may be present at that location. Val/Leu/Phe/Ile Asn/Asp/Glu/Ser Leu/ILe/Phe/Val/Gly Thr/Ser/Ala/Gly 486 Thr/His/Asp/Asn Ser/Asn/Asp/Ile Asp/Glu Leu/Met Val/Ala Lys/Val/Ala 490                                                     495 Asp/Ile/Glu/Ser His Leu/Gly/Ile/Val Arg/Lys Tyr/His Leu/Met Met/Val                                             500 Gly Gly     504

[0055] It is understood that analogous substitutions throughout the sequence are encompassed within the scope of the invention, and that Region R4 is simply used above for purposes of illustration.

[0056] Another manner in which similarity may exist between two amino acid sequences is where a given amino acid is substituted with another amino acid from the same amino acid group. In this manner, it is known that serine may commonly be substituted with threonine in a polypeptide without substantially altering the functionality of the polypeptide. The following sets forth groups of amino acids which are believed to be interchangeable in inventive amino acid sequences at a wide variety of locations without substantially altering the functionality thereof: Group I: Nonpolar amino acids: Alanine, valine, proline, leucine, phenylalanine, tryptophan, methionine, isoleucine, cysteine, glycine; Group II: Uncharged polar amino acids: Serine, threonine, asparagine, glutamine, tyrosine; Group III: Charged polar acidic amino acids: Aspartic, glutamic; and Group IV: Charged polar basic amino acids: Lysine, arginine, histidine.

[0057] Where one is unsure whether a given substitution will affect the functionality of the enzyme, this may be determined without undue experimentation using synthesis techniques and screening assays known in the art.

[0058] Having established the meaning of similarity with respect to an amino acid sequence, it is important to note that the invention features mutated amino acid sequences comprising one or more amino acid substitutions that do alter the functionality of the wild-type TD enzyme. Inventive insensitive TD enzymes are therefore not similar to wild-type TD, as that term is defined and used herein, because inhibition functionality is altered. Insensitive TD enzymes feature one or more mutations in the regulatory site which mutations alter the functionality of the regulatory site without substantially altering the functionality of the catalytic site. In one specific aspect of the invention, there is provided an amino acid sequence (SEQ ID NO:2) having two substitutions, this sequence comprising a mutated TD which has good catalytic functionality but which does not exhibit regulatory functionality. In other words, the enzyme set forth in SEQ ID NO:2 comprises a feedback insensitive Arabidopsis thaliana TD.

[0059] It is seen upon comparing the wild type TD set forth in SEQ ID NO:1 and the mutated sequence of SEQ ID NO:2, which comprises a specific embodiment of the invention, that the sequences differ only by two point mutations in the respective nucleotide sequences (C to T at nucleotide 1495; and G to A at nucleotide 1631), which result in two amino acid substitutions in the TD polypeptide (Arg to Cys at amino acid location 499; and Arg to His at amino acid location 544). The first mutation is in regulatory region R4 of TD, and the second is in regulatory region R6 of TD. The Arg to Cys substitution at amino acid residue 499 changed a charged, polar, basic amino acid (Arg) to a nonpolar amino acid (Cys) which altered the feedback site in TD. On the other hand, the change of Arg to His at residue 544 was a change from a charged, polar, basic amino acid (Arg) to another charged, polar, basic amino acid (His). While it is not intended that the present invention be limited by any theory by which it achieves its advantageous result, it is believed that the substitution at residue 544 alone may not have substantially altered the feedback site of TD, and, in contrast, that the substitution at residue 499 alone may have desensitized TD encoded thereby to feedback regulation. Certainly, when combined, the substitutions were very effective in desensitizing TD encoded by omr1 to feedback regulation.

[0060] It is recognized that the amino acid sequence set forth in SEQ ID NO:3 (585 residues encoded by omr1) is a truncated version, missing 7 amino-terminal residues, of that set forth in SEQ ID NO:2. It is seen from the following description, including the Examples set forth herein, that a significant amount of research was performed based upon this slightly shortened version, and that the slightly shortened version may be advantageously used to transform a wide variety of plants and microorganisms. It is believed that the portion of the amino acid sequence that is present in SEQ ID NO:2 and absent in SEQ ID NO:3 is a portion of the chloroplast leader sequence, and not present in the mature TD enzyme.

[0061] As mentioned above, to assist in the description of the present invention, SEQ ID NO:1 is provided which sets forth a nucleotide sequence, and the amino acid sequence encoded thereby, comprising a wild-type TD from Arabidopsis thaliana. SEQ ID NOS:2 and 3 set forth nucleotide sequences, and amino acid sequences encoded thereby, comprising precursor proteins of differing lengths. SEQ ID NO:3 (see also FIG. 6b) encodes a 609 amino acid fusion or chimeric polypeptide of which 585 amino acid residues are encoded by mutant omr1 of Arabidopsis. That is, SEQ ID NO:3 encodes a mutant TD that is shorter than the full-length mutant TD shown in SEQ ID NO:2 by 7 amino terminal residues. Since transgenic plants transformed with pCM35s-omr1 were capable of expressing OMT resistance, then the 585 amino acid-long truncated precursor was fully capable of translocation from the cytoplasm to the chloroplast. SEQ ID NOS:4, 5 and 6 set forth sequences comprising three predicted mature proteins. SEQ ID NO:7 sets forth the putative regulatory site of an inventive mutated TD enzyme, and SEQ ID NOS:8 and 9 set forth regulatory regions harboring mutations in accordance with one aspect of the invention.

[0062] It is understood that the wild-type TD enzyme features dual functionality. Specifically, the TD enzyme has a catalytic site which is divided into catalytic regions C1-C5, as shown with respect to the analogous tomato TD enzyme and chickpea TD enzyme in FIG. 2. The catalytic site catalyzes the reaction of threonine to 2-oxobutyrate. TD also has a regulatory site which is divided into regulatory regions R1-R7, as shown in FIG. 2. The regulatory site is responsible for the feedback inhibition which occurs when the regulatory site binds to an inhibitor, in this case isoleucine.

[0063] The present application finds advantageous use in a wide variety of plants, as well as in a wide variety of microorganisms. With respect to plants, it is important to recognize that the TD enzyme functions in chloroplasts, and, therefore, that the polypeptide transcribed therefore is a precursor protein which includes a portion identified herein as a “chloroplast leader sequence.” For purposes of the present description, the term “chloroplast leader sequence” is used interchangeably with the term “transit peptide.” The chloroplast leader sequence is covalently bound to the “mature enzyme” or “passenger enzyme.” The term “precursor protein” is meant a polypeptide having a transit peptide and a passenger peptide covalently attached to each other. Typically, the carboxy terminus of the transit peptide is covalently attached to the amino terminus of the passenger peptide. The passenger peptide and transit peptide can be encoded by the same gene locus, that is, homologous to each other, in that they are encoded in a manner isolated from a single source. Alternatively, the transit peptide and passenger peptide can be heterologous to each other, i.e., the transit peptide and passenger peptide can be from different genes and/or different organisms. The terms “transit peptide,” “chloroplast leader sequence,” and “signal peptide” are used interchangeably to designate those amino acids that direct a passenger peptide to a chloroplast. By “mature peptide” or “passenger peptide” is meant a polypeptide which is found after processing and passing into an organelle and which is functional in the organelle for its intended purpose. Passenger peptides are originally made in a precursor form that includes a transit peptide and the passenger peptide. Upon entry into an organelle, the transit peptide portion is cleaved, thus leaving the “passenger” or “mature” peptide. Passenger peptides are the polypeptides typically obtained upon purification from a homogenate, the sequence of which can be determined as described herein.

[0064] The transit peptide may be derived from monocotyledonous or dicotyledonous plants upon choice of the artisan. DNA sequences encoding said transit peptides may be obtained from chloroplast proteins such as Δ-9 desaturase, palmitoyl-ACP thioesterase, β-KETOACYL-ACP synthase, oleyl-ACP thioesterase, chlorophyll a/b binding protein, NADPH+ dependent glyceraldehyde-3-phosphate dehydrogenase, early light inducible protein, clip protease regulatory protease, pyruvate orthophosphate dikinase, chlorophyll a/b binding protein, triose phosphate3-pohosphoglycerate phosphate translocator, 5-enol pyruval shikimate-e-phosphate synthase, dihydrofolate reductase, thymidylate synthase, acetyl-coenzyme A carboxylase, Cu/Zn superoxide dismutase, cystein synthase, rubisco activase, ferritin, granule bound starch synthase, pyrophosphate, glutamine synthase, aldolase, glutathione reductase, nitrite reductase, 2-oxoglutarate/malate translocator, ADP-glucose pyrophosphorylase, ferrodoxin, carbonic anhydrase, polyphenol oxidase, ferrodoxin NADP=oxidoreductase, platocyannin, glycerol-3-phosphate dehydrogenase, lipoxygenase, o-acetylserine (thiol)-lysase, acyl carrier protein, 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase, chloroplast-localized heat shock protein, starch phosphorylase, pyruvate orthophosphate dikinase, starch glycosyltrtansferase, and the like, of which the transit peptide portion has been defined in GenBank.

[0065] In plants, the chloroplast leader sequence is used to direct the passenger protein to chloroplasts; however, they are typically cleaved and degraded upon entry of the passenger protein into the organelle of interest. Therefore, purification of a cleaved transit peptide from plant tissues is typically not possible. In some cases, however, transit peptide sequences can be determined by comparison of the precursor protein amino acid sequence obtained from the gene encoding the same to the amino acid sequence of the isolated passenger protein (mature protein). Furthermore, passenger protein sequences can also be determined from the transit peptide proteins associated therewith by comparison of sequences to other similar proteins isolated from different species. As exemplified herein, genes encoding precursor forms of mutated TD protein, disclosed as SEQ ID NO:2 and SEQ ID NO:3, when compared to wild type precursor and mature TD protein obtained from other species, can establish the expected sequence of the mature protein.

[0066] As previously discussed, the amino acid sequence and hence the nucleic acid sequence of a transit peptide can be determined in a variety of ways available to the skilled artisan. For example, passenger proteins of interest can be purified using a variety of techniques available to the person skilled in the art of protein biochemistry. Once purified, an amino terminal sequence of the protein can be determined using methods such as Edman degradation, mass spectroscopy, nuclear magnetic spectroscopy and the like. Using this information and the genetic code, standard molecular biology techniques can be employed to clone the gene encoding the protein as exemplified herein. Comparison of amino acid sequence determined from the cDNA to that obtained from the amino terminal sequence of the passenger protein can allow determination of the transit peptide sequence. In addition, many transit peptide sequences are available in the art and can easily be obtained form GenBank located in the Entrez Database at the National Center for Biotechnology Information web site.

[0067] The subject of transit peptides in plants has been extensively reviewed by Keegstra et al., (1989) (Cell, 56:247-253), which is incorporated herein by reference. Typically, there is very little primary amino acid sequence homology between different plant transit peptides. Even though passenger proteins may have amino acid and nucleic acid sequence similarities between cultivars, lines, and species, transit peptide may show very little sequence homology at any level. Furthermore, the length of transit peptides can vary, with some precursor proteins comprising transit peptide proteins with as few as about 10 amino acids while others can be about 150 amino acids or longer. Additional descriptions of transit peptide characteristics in plants and mechanisms associated therewith can be found in Ko and Ko, (1992) J. Biol. Chem. 267, 13910-13916; Bascomb et al. (1992) Plant Microb. Biotechnol. Res. Ser. 1:142-163; and Bakau et al., (1996) Trends in Cell Biol. 6:480-486; which are incorporated herein by reference.

[0068] In this regard, the first 90 amino acid residues in the N-terminal region of the Arabidopsis TD protein encoded by omr1 (in SEQ ID NO:2) represent an expected region comprising the transit peptide, as indicated by:

[0069] (i) the dissimilarity with the yeast, Salmonella and E. coli TD proteins,

[0070] (ii) the comparison of the sizes of TD of Arabidopsis, tomato, chickpea, yeast, Salmonella and E. coli, and

[0071] (iii) the amino acid composition which contains 12 proline residues and 33 other hydrophobic residues constituting a total of 50% hydrophobic residues.

[0072] Therefore, it is expected that the mature/passenger TD of Arabidopsis encoded by the omr1 locus, cleavage of the transit peptide may occur at the peptide bond between the alanine at residue 90 and the glutamic acid at residue 91, leaving behind a mature/passenger TD that starts at the glutamic acid at residue 91. As such, SEQ ID NO:4 identifies an expected mature TD for Arabidopsis that starts at the glutamic acid at residue 91 of SEQ ID NO:2 (clone 592). This expected mature TD polypeptide comprises 502 sequential amino acid residues.

[0073] The only two other higher plant TD genes that have been cloned to date are those of tomato (Samach A., Harven D., Gutfinger T., Ken-Dror S., Lifschitz E., 1991, Proc Natl Acad Sci USA 88:2678-2682) and chickpea (Jacob John S., Srivastava V., Guha-Mukherjee S., 1995, Plant Physiol 107:1023-1024). The lengths of the transit peptides of the tomato TD and chickpea TD were predicted to be the first −80 and 91 amino terminal residues, respectively, and the full length precursor proteins were reported to be 595 residues and 590 residues, respectively (Samach et al., 1991; Jacob John et al., 1995). In both tomato and chickpea, the amino-terminus of the TD protein contained a typical two-domain transit peptide consistent with chloroplast lumen targeting sequences (Keegstra K., Olsen L. J., Theg S. M., 1989, Chloroplast precursors and their transport across the membrane. Annu Rev Plant Physiol Plant Mol Biol 40:471-501). In tomato, the first domain at the amino-terminal (45 residues) of the transit peptide was rich in serine and threonine (33%) while the following sequence of 35 residues contained 8 regularly spaced proline and other hydrophobic residues (Samach et al., 1991). By sequencing the first ten amino-terminal residues of a purified tomato TD from flowers, Samach et al., (1991) found that lysine at residue 52 is the first amino acid at the amino-terminal end of the mature/passenger protein. According to Samach et al., (1991), the hydrophobic domain of the transit peptide of tomato TD is not cleaved and remains as part of the mature TD in the chloroplast. Samach et al., (1991) also explained that “it is possible that only a fraction of the tomato TD protein is cleaved at position 52, while the rest of the transit peptide is cleaved elsewhere and remain refractory to amino-terminal sequencing.” In chickpea, the first domain at the amino-terminal end of the transit peptide was deduced to be 45 residues and rich in threonine and serine (37%) while the remaining 46 residues contained 8 regularly spaced proline residues and 19 other hydrophobic residues (Jacob John et al., 1995). The cleavage site of the transit peptide of chickpea TD was not determined.

[0074] By analogy to tomato and chickpea, Arabidopsis TD also showed a typical two-domain transit peptide consistent with chloroplast lumen targeting sequences (as reviewed by Keegstra et al., 1989). The first 49 residues of the amino terminal end represented a domain that was rich in serine and threonine (31%) and other hydrophilic residues while the remaining 41 residues represented a second domain that contained 59% hydrophobic residues. The cleavage site of the transit peptide of Arabidopsis TD was not determined. Therefore, by analogy to tomato, it is expected that the cleavage site of the transit peptide of Arabidopsis TD may alternatively start at the lysine at residue 54 or at the lysine at residue 61. This is a presumptive cleavage site and one skilled in the art can readily determine the cleavage site in a similar fashion as in the case of tomato (Samach et al., 1991) by purifying Arabidopsis TD then sequencing the first ten amino acids in the amino-terminal end. Therefore, two additional sequences are provided as SEQ ID NOS:5 and 6 that alternatively identify two expected mature TD in Arabidopsis.

[0075] It is within the scope of the present invention to create chimeric polynucleotides encoding precursor proteins wherein a transit peptide of choice is in the proper reading frame with the mature coding sequence of mutated TD. As used herein, the terms “chimeric polynucleotide,” “chimeric DNA construct” and “chimeric DNA” are used to refer to recombinant DNA.

[0076] In creating a chimeric DNA construct encoding a transit peptide as disclosed herein, the transit peptide being heterologous to the mature, mutated TD, the DNA encoding the transit peptide is place 5′ and in the proper reading frame with the DNA encoding the mature, mutated TD protein. Placement of the chimeric DNA in correct relationship with promoter regulatory elements and other sequences as described herein can allow production of mRNA molecules that encode for heterologous precursor proteins. By “promoter regulatory element” is meant nucleotide sequence elements within a nucleotide sequence which control the expression of that nucleotide sequence. Promoter regulatory elements provide the nucleic acid sequences necessary for recognition of RNA polymerase and other transcriptional factors required for efficient transcription. Promoter regulatory elements are meant to include constitutive, tissue-specific, developmental-specific, inducible promoters and the like. Promoter regulatory elements may also include certain enhancer sequence elements that improve transcriptional efficiency. The mRNA can then be translated thus producing a functional heterologous precursor protein which can be delivered to the chloroplast. It is, of course, understood that a DNA construct may be made in accordance with the invention to include a promotor that is native to the gene of a selected species that encodes that species' TD precursor polypeptide. Uptake of the protein by the chloroplast and cleavage of the associated transit peptide can result in a chloroplast containing a mature, mutated form of TD, thus rendering the cell resistant to feedback inhibition which would normally inhibit cells containing only the wild-type TD protein.

[0077] The present invention, therefore, provides, in alternative aspects, a feedback insensitive TD comprising the amino acid sequence set forth in SEQ ID NO:2 or SEQ ID NO:3 (precursor polypeptides); set forth in SEQ ID NO:4, SEQ ID NO:5 or SEQ ID NO:6 (expected mature TD enzymes); SEQ ID NO:7 (an insensitive TD regulatory site); and set forth in SEQ ID NO:8 (regulatory region R4) or SEQ ID NO:9 (regulatory region R6). SEQ ID NO:7 or variants thereof as described above, may be operably coupled to a sequence encoding a TD catalytic site from a wide variety of species, including functionally similar variants thereof, to provide the advantageous result of the invention.

[0078] It is readily understood that, in the case of transforming prokaryotes, it is not necessary to include a transit peptide in the coding region of the vector. Rather, since such cells do not possess chloroplasts, an inventive DNA construct for transforming, for example, bacteria, may be made by simply attaching a start codon directly to, and in the proper reading frame with, a mature peptide. Of course, other elements are preferably present as described herein, such as a promoter upstream of the start codon and a termination sequence downstream of the coding region.

[0079] SEQ ID NOS:8 and 9 may also be operably coupled to a wide variety of sequences to provide insensitive TD enzymes, and therefore comprise certain preferred aspects of the invention. Substitutions giving rise to similar amino acid sequences, as described herein, are particularly applicable to SEQ ID NO:8, and the following sets forth a plurality of particularly preferred alternative sequences for SEQ ID NO:8 in accordance with the invention: Val/Leu/Phe/Ile Asn/Asp/Glu/Ser Leu/Ile/Phe/Val/Gly Thr/Ser/Ala/Gly Thr/His/Asp/ASn Ser/Asn/Asp/Ile Asp/Glu Leu/Met Val/Ala Lys/Val/Ala Asp/Ile/Glu/Ser His Leu/Gly/Ile/Val Cys Tyr/His Leu/Met Met/Val Gly Gly

[0080] The invention therefore also encompasses amino acid sequences similar to the amino acid sequences set forth herein that have at least about 50% identity thereto and that are insensitive to feedback inhibition by Ile. Preferably, inventive amino acid sequences have at least about 75% identity to these sequences, more preferably at least about 85% identity and most preferably at least about 95% identity.

[0081] Percent identity may be determined, for example, by comparing sequence information using the GAP computer program, version 6.0, available from the University of Wisconsin Genetics Computer Group (UWGCG). The GAP program utilizes the alignment method of Needleman and Wunsch (J. Mol. Biol. 48:443, 1970), as revised by Smith and Waterman (Adv. Appl. Math. 2:482, 1981). Briefly, the GAP program defines identity as the number of aligned symbols (i.e., nucleotides or amino acids) which are the same, divided by the total number of symbols in the shorter of the two sequences. The preferred default parameters for the GAP program include: (1) a uniary comparison matrix (containing a value of 1 for identities and 0 for non-identities), and the weighted comparison matrix of Gribskov and Burgess, Nucl. Acids Res. 14:6745, 1986, as described by Schwartz and Dayhoff, eds., Atlas of protein Sequence and Structure, National Biomedical Research Foundation, pp. 353-358, 1979; (2) a penalty of 3.0 for each gap and an additional 0.10 penalty for each symbol in each gap; and (3) no penalty for end gaps.

[0082] The invention also contemplates amino acid sequences having alternative mutations to those identified herein which also result in a feedback insensitive TD. For example, it is expected that the cys at position 499 and the his at position 544 in SEQ ID NO:2 could be substituted with alternative amino acids from the same amino acid group as cys and his, respectively (as described above) to provide an alternate inventive enzyme. Further, it is well within the purview of a person skilled in the art to engineer a feedback insensitive TD by providing a wild-type TD and substituting a highly conserved amino acid at a given location in the regulatory site with a diverse amino acid (i.e., one from a different amino acid group), and to assay the resulting enzyme for catalytic activity and feedback sensitivity. For example, a skilled artisan can alter the nucleotide sequence set forth in SEQ ID NO:1 by site-directed mutagenesis to provide a mutated sequence which encodes an enzyme having an alternate amino acid in a given location of the enzyme. Alternatively, a skilled artisan can synthesize an amino acid sequence having one or more additions, substitutions and/or deletions at a highly conserved location of the wild-type TD enzyme using techniques known in the art. Such variants, which exhibit functionality substantially similar to a polypeptide comprising the sequence set forth in SEQ ID NO:2, are included within the scope of the present invention.

[0083] Turning now to nucleotide sequences encoding inventive insensitive TD enzymes, nucleotide sequences encoding preferred feedback insensitive precursor TD of the species Arabidopsis thaliana are set forth in SEQ ID NOS:2 and 3 herein. The mutated polynucleotides set forth therein are referred to as omr1. omr1 has been found to be a dominant allele, this imparting significant value to the invention. It is of course not intended that the present invention be limited to this exemplary nucleotide sequence, but include sequences having substantial identity thereto and sequences which encode variant forms of insensitive TD as described above.

[0084] The term “nucleotide sequence,” as used herein, is intended to refer to a natural or synthetic linear and sequential array of nucleotides and/or nucleosides, and derivatives thereof. The terms “encoding” and “coding” refer to the process by which a nucleotide sequence, through the mechanisms of transcription and translation, provides the information to a cell from which a series of amino acids can be assembled into a specific amino acid sequence to produce a functional polypeptide, such as, for example, an active enzyme. The process of encoding a specific amino acid sequence may involve DNA sequences having one or more base changes (i.e., insertions, deletions, substitutions) that do not cause a change in the encoded amino acid, or which involve base changes which may alter one or more amino acids, but do not eliminate the functional properties of the polypeptide encoded by the DNA sequence.

[0085] It is therefore understood that the invention encompasses more than the specific exemplary nucleotide sequence of omr1. For example, a nucleic-acid sequence encoding a variant amino acid sequence, as discussed above, is within the scope of the invention. Modifications to a sequence, such as deletions, insertions, or substitutions in the sequence which produce “silent” changes that do not substantially affect the functional properties of the resulting polypeptide molecule are expressly contemplated by the present invention. For example, it is understood that alterations in a nucleotide sequence which reflect the degeneracy of the genetic code, or which result in the production of a chemically equivalent amino acid at a given site, are contemplated. Thus, a codon for the amino acid alanine, a hydrophobic amino acid, may be substituted by a codon encoding another less hydrophobic residue, such as glycine, or a more hydrophobic residue, such as valine, leucine, or isoleucine. Similarly, changes which result in substitution of one negatively charged residue for another, such as aspartic acid for glutamic acid, or one positively charged residue for another, such as lysine for arginine, can also be expected to produce a biologically equivalent product.

[0086] Nucleotide changes which result in alteration of the N-terminal and C-terminal portions of the polypeptide molecule would also not be expected to alter the activity of the polypeptide. In some cases, it may in fact be desirable to make mutations in the sequence in order to study the effect of alteration on the biological activity of the polypeptide. Each of the proposed modifications is well within the routine skill in the art.

[0087] In a preferred aspect, therefore, the present invention contemplates nucleotide sequences having substantial identity to the sequences set forth herein and variants thereof as described herein. The term “substantial identity” is used herein with respect to a nucleotide sequence to designate that the nucleotide sequence has a sequence sufficiently similar to a reference nucleotide sequence that it will hybridize therewith under moderately stringent conditions, this method of determining identity being well known in the art to which the invention pertains. Briefly, moderately stringent conditions are defined in Sambrook et al., Molecular Cloning: a Laboratory Manual, 2ed. Vol. 1, pp. 101-104, Cold Spring Harbor Laboratory Press (1989) as including the use of a prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0) and hybridization and washing conditions of about 55° C., 5×SSC. A further requirement of an inventive polynucleotide variant is that it must encode a polypeptide having similar functionality to the specific mutated TD enzymes recited herein, i.e., good catalytic functionality and insensitivity to feedback inhibition.

[0088] A suitable DNA sequence selected for use according to the invention may be obtained, for example, by cloning techniques using cDNA libraries corresponding to a wide variety of species, these techniques being well known in the relevant art. Suitable nucleotide sequences may be isolated from DNA libraries obtained from a wide variety of species by means of nucleic acid hybridization or PCR, using as hybridization probes or primers nucleotide sequences selected in accordance with the invention, such as those set forth in SEQ ID NOS:1-10; nucleotide sequences having substantial identity thereto; or portions thereof. Isolated wild-type sequences encoding TD may then be altered as provided by the present invention by site-directed mutagenesis.

[0089] Alternatively, a suitable sequence may be made by techniques which are also well known in the art. For example, nucleic acid sequences encoding enzymes of the invention may be constructed using standard recombinant DNA technology, for example, by cutting or splicing nucleic acids which encode cytokines and/or other peptides using restriction enzymes and DNA ligase. Alternatively, nucleic acid sequences may be constructed using chemical synthesis, such as solid-phase phosphoramidate technology. In preferred embodiments of the invention, polymerase chain reaction (PCR) is used to accomplish splicing of nucleic acid sequences by overlap extension as is known in the art.

[0090] Inventive DNA sequences can be incorporated into the genome of a plant or microorganism using conventional recombinant DNA technology, thereby making a transformed plant or microorganism having the excellent features described herein. In this regard, the term “genome” as used herein is intended to refer to DNA which is present in a plant or microorganism and which is heritable by progeny during propagation thereof. As such, an inventive transformed plant or microorganism may alternatively be produced by producing F1 or higher generation progeny of a directly transformed plant or microorganism, wherein the progeny comprise the foreign nucleotide sequence. Transformed plants or microorganisms and progeny thereof are all contemplated by the invention and are all intended to fall directly within the meaning of the terms “transformed plant” and “transformed microorganism.”

[0091] In this manner, the present invention contemplates the use of transformed plants which are selfed to produce an inbred plant. The inbred plant produces seed containing the gene of interest. These seeds can be grown to produce plants that express the protein of interest. The inbred lines can also be crossed with other inbred lines to produce hybrids. Parts obtained from the regenerated plant, such as flowers, seeds, leaves, branches, fruit, and the like are covered by the invention provided that said parts contain genes encoding and/or expressing the protein of interest. Progeny and variants, and mutants of the regenerated plants are also included within the scope of the invention.

[0092] In diploid plants, typically one parent may be transformed and the other parent is the wild type. After crossing the parents, the first generation hybrids (F1) are selfed to produce second generation hybrids (F2). Those plants exhibiting the highest levels of the expression can then be chosen for further breeding.

[0093] Genes encoding precursor mutated TD polypeptides, as disclosed herein as SEQ ID NO:2 and SEQ ID NO:3, can be used in conjunction with other plant regulatory elements to create plant cells expressing the polypeptides. By “expressing” as used herein, is meant the transcription and stable accumulation of mRNA inside a cell, the cell being of prokaryotic or eukaryotic origin. Furthermore, it is within the scope of the invention to place mutated mature TD from Arabidopsis into other species including monocotyledonous and dicotyledonous plants. In so doing, chimeric gene constructs encoding the mature, mutated TD proteins having transit peptides heterologous thereto (transit peptides from a different protein or species) can be used. Transit peptides of the present invention, when covalently attached to the mature, mutated TD protein, can provide intracellular transport to the chloroplast. In plants, a mutated mature form of TD found in a chloroplast of a cell renders the cell resistant to feedback inhibition and resistance to Ile structural analogs.

[0094] Generally, transformation of a plant or microorganism involves inserting a DNA sequence into an expression vector in proper orientation and correct reading frame. The vector may desirably contain the necessary elements for the transcription of the inserted polypeptide-encoding sequence. A wide variety of vector systems known in the art can be advantageously used in accordance with the invention, such as plasmids, bacteriophage viruses or other modified viruses. Suitable vectors include, but are not limited to the following viral vectors: lambda vector system gt11, gt10, Charon 4, and plasmid vectors such as pBI121, pBR322, pACYC177, pACYC184, pAR series, pKK223-3, pUC8, pUC9, pUC18, pUC19, pLG339, pRK290, pKC37, pKC101, pcDNAII, and other similar systems. The DNA sequences may be cloned into the vector using standard cloning procedures in the art, for example, as described by Maniatis et al., Molecular Cloning: A Laboratory Manual, Cold Springs Laboratory, Cold Springs Harbor, N.Y. (1982), which is hereby incorporated by reference in its entirety. The plasmid pBI121 is available from Clontech Laboratories, Palo Alto, Calif. It is understood that known techniques may be advantageously used according to the invention to transform microorganisms such as, for example, Agrobacterium sp., yeast, E. coli and Pseudomonas sp.

[0095] In order to obtain satisfactory expression of a nucleotide sequence which encodes an inventive feedback insensitive TD in a plant or microorganism, it is preferred that a promoter be present in the expression vector. The promoter is preferably a constitutive promoter, but may alternatively be a tissue-specific promoter or an inducible promoter. Preferably, the promoter is one isolated from a native gene which encodes a TD. Although promoters for certain classes of genes commonly differ between species, it is understood that the present invention includes promoters which regulate expression of a wide variety of genes in a wide variety of plant or microorganism species.

[0096] An expression vector according to the invention may be either naturally or artificially produced from parts derived from heterologous sources, which parts may be naturally occurring or chemically synthesized, and wherein the parts have been joined by ligation or other means known in the art. The introduced coding sequence is preferably under control of the promoter and thus will be generally downstream from the promoter. Stated alternatively, the promoter sequence will be generally upstream (i.e., at the 5′ end) of the coding sequence. The phrase “under control of” contemplates the presence of such other elements as may be necessary to achieve transcription of the introduced sequence. As such, in one representative example, enhanced production of a feedback insensitive TD may be achieved by inserting an inventive nucleotide sequence in a vector downstream from and operably linked to a promoter sequence capable of driving expression in a host cell. Two DNA sequences (such as a promoter region sequence and a feedback insensitive TD-encoding nucleotide sequence) are said to be operably linked if the nature of the linkage between the two DNA sequences does not (1) result in the introduction of a frame-shift mutation, (2) interfere with the ability of the promoter region sequence to direct the transcription of the desired nucleotide sequence, or (3) interfere with the ability of the desired nucleotide sequence to be transcribed by the promoter region sequence.

[0097] RNA polymerase normally binds to the promoter and initiates transcription of a DNA sequence or a group of linked DNA sequences and regulatory elements (operon). A transgene, such as a nucleotide sequence selected in accordance with the present invention, is expressed in a transformed cell to produce in the cell a polypeptide encoded thereby. Briefly, transcription of the DNA sequence is initiated by the binding of RNA polymerase to the DNA sequence's promoter region. During transcription, movement of the RNA polymerase along the DNA sequence forms messenger RNA (“mRNA”) and, as a result, the DNA sequence is transcribed into a corresponding mRNA. This mRNA then moves to the ribosomes of the cytoplasm or rough endoplasmic reticulum which, with transfer RNA (“tRNA”), translates the mRNA into the polypeptide encoded thereby.

[0098] It is well known that there may or may not be other regulatory elements (e.g., enhancer sequences) which cooperate with the promoter and a transcriptional start site to achieve transcription of the introduced (i.e., foreign) coding sequence. By “enhancer” is meant nucleotide sequence elements which can stimulate promoter activity in a cell such as those found in plants as exemplified by the leader sequence of maize streak virus (MSV), alcohol dehydrogenase intron 1, and the like. Also, the recombinant DNA will preferably include a transcriptional termination sequence downstream from the introduced sequence. It may also be desirous to use a reporter gene. In some instances, a reporter gene may be used with or without a selectable marker. Reporter genes are genes which are typically not present in the recipient organism or tissue and typically encode proteins resulting in some phenotypic change or enzymatic property. Examples of such genes are provided in K. Wising et al. (1988) Ann. Rev. Genetics, 22:421, which is incorporated herein by reference. Preferred reporter genes include the beta-glucuronidase (GUS) of the uidA locus of E. coli, the green fluorescent protein from the bioluminescent jellyfish Aequorea victoria, and the luciferase genes from firefly Photinus pyralis. An assay for detecting reporter gene expression may then be performed at a suitable time after the gene has been introduced into recipient cells. A preferred such assay entails the use of the gene encoding beta-glucuronidase (GUS) of the uidA locus of E. coli, as described by Jefferson et al., (1987 Biochem. Soc. Trans. 15, 17-19) to identify transformed cells.

[0099] Plant promoter regulatory elements from a wide variety of sources can be used efficiently in plant cells to express foreign genes. For example, promoter regulatory elements of bacterial origin, such as the octopine synthase promoter, the nopaline synthase promoter, the mannopine synthase promoter, and promoters of viral origin, such as the cauliflower mosaic virus (35S and 19S), 35T (which is a re-engineered 35S promoter, WO 97/13402 published Apr. 17, 1997) and the like may be used. Plant promoter regulatory elements include, but are not limited to, ribulose-1-5-bisphosphate (RUBP) carboxylase small subunit (ssu), beta-conglycinin promoter, beta-phaseolin promoter, ADH promoter, heat-shock promoters, and tissue-specific promoters.

[0100] Other elements such as matrix attachment regions, scaffold attachment regions, introns, enhancers, polyadenylation sequences, and the like, may be present and thus may improve the transcription efficiency or DNA integration. Such elements may or may not be necessary for DNA function, although they can provide better expression or functioning of the DNA by affecting transcription, mRNA stability, and the like. Such elements may be included in the DNA as desired to obtain optimal performance of the transformed DNA in the plant. Typical elements include, but are not limited to, Adh-intron 1, Adh-intron 0.6, the alfalfa mosaic virus coat protein leader sequence, the maize streak virus coat protein leader sequence, as well as others available to a skilled artisan.

[0101] Constitutive promoter regulatory elements may be used thereby directing continuous gene expression in all cell types at all times (e.g., actin, ubiquitin, CaMV 35S, and the like). Tissue specific promoter regulatory elements are responsible for gene expression in specific cell or tissue types, such as the leaves or seeds (e.g., zein, oleosin, napin, ACP, globulin, and the like) and these may alternatively be used.

[0102] Promoter regulatory elements may also be active during a certain stage of the plants' development as well as active in plant tissues and organs. Examples of such include, but are not limited to, pollen-specific, embryo-specific, corn silk-specific, cotton fiber-specific, root-specific, seed endosperm-specific promoter regulatory elements, and the like. Under certain circumstances, it may be desirable to use an inducible promoter regulatory element, which is responsible for expression of genes in response to a specific signal, such as, for example, physical stimulus (heat shock genes), light (RUBP carboxylase), hormone (Em), metabolites, chemicals and stress Other desirable transcription and translation elements that function in plants may also be used. Numerous plant-specific gene transfer vectors are known in the art.

[0103] Once the DNA construct of the present invention has been cloned into an expression vector, it may then be transformed into a host cell. In addition to numerous technologies for transforming plants, the type of tissue which is contacted with foreign polynucleotides may vary as well. Plant tissue suitable for transformation of a plant in accordance with certain preferred aspects of the invention include, for example, whole plants, leaf tissues, flower buds, root tissues, callus tissue types I, II and III, embryogenic tissue, meristems, protoplasts, hypocotyls and cotyledons. It is understood, however, that this list is not intended to be limiting, but only to provide examples of plant tissues which may be advantageously transformed in accordance with the present invention. A wide variety of plant tissues may be transformed during dedifferentiation using appropriate techniques described herein.

[0104] Transformation of a plant or microorganism may be achieved using one of a wide variety of techniques known in the art. The manner in which the transcriptional unit is introduced into the plant host is not critical to the invention. Any method which provides efficient transformation may be employed. One technique of transforming plants with a DNA construct inaccordance with the present invention is by contacting the tissue of such plants with an inoculum of bacteria transformed with a vector comprising the DNA construct. Generally, this procedure involves inoculating the plant tissue with a suspension of bacteria and incubating the tissue for about 48 to about 72 hours on regeneration medium without antibiotics at about 25-28° C. Bacteria from the genus Agrobacterium may be advantageously utilized to transform plant cells. Suitable species of such bacterium include Agrobacterium tumefaciens and Agrobacterium rhizogenes. Agrobacterium tumefaciens (e.g., strains LBA4404 or EHA105) is particularly useful due to its well-known ability to transform plants. Another technique which may advantageously be used is vacuum-infiltration of flower buds using Agrobacterium-based vectors.

[0105] Various methods for plant transformation include the use of Ti or Ri-plasmids and the like to perform Agrobacterium mediated transformation. In many instances, it will be desirable to have the construct used for transformation bordered on one or both sides by T-DNA borders, more specifically the right border. This is particularly useful when the construct uses Agrobacterium tumefaciens or Agrobacterium rhizogenes as a mode for transformation, although T-DNA borders may find used with other modes of transformation. Where Agrobacterium is used for plant transformation, a vector may be used which may be introduced into the host for homologous recombination with T-DNA or the Ti or Ri plasmid present in the host. Introduction of the vector may be performed via electroporation, tri-parental mating and other techniques for transforming gram-negative bacteria which are known to those skilled in the art. The manner of vector transformation into the Agrobacterium host is not critical to the invention.

[0106] In some cases where Agrobacterium is used for transformation, the expression construct being within the T-DNA borders will be inserted into a broad spectrum vector such as pRK2 or derivatives thereof as described in Ditta et al. (PNAS USA (1980) 77:7347-7351 and EPO 0 120 515), which are incorporated herein by reference. Explants may be combined and incubated with the transformed Agrobacterium for sufficient time to allow transformation thereof. After transformation, the Agrobacteria and plant cells are cultured with the appropriate selective medium. Once calli are formed, shoot formation can be encouraged by employing the appropriate plant hormones according to methods well known in the art of plant tissue culturing and plant regeneration. However, a callus intermediate stage is not always-necessary. After shoot formation, said plant cells can be transferred to medium which encourages root formation thereby completing plant regeneration. The plants may then be grown to seed and the seed can be used to establish future generations. Regardless of transformation technique, the polynucleotide of interest is preferably incorporated into a transfer vector adapted to express the polynucleotide in a plant cell by including in the vector a plant promoter regulatory element, as well as 3′ non-translated transcriptional termination regions such as Nos and the like.

[0107] Plant RNA viral based systems can also be used to express genes for the purposes disclosed herein. In so doing, the chimeric genes of interest can be inserted into the coat promoter regions of a suitable plant virus under the control of a subgenomic promoter which will infect the host plant of interest. Plant RNA viral based systems are described, for example, in U.S. Pat. Nos. 5,500,360; 5,316,931 and 5,589,367, each of which is hereby incorporated herein by reference in its entirety.

[0108] Another approach to transforming plant cells with a DNA sequence selected in accordance with the present invention involves propelling inert or biologically active particles at plant tissues or cells. This technique is disclosed in U.S. Pat. Nos. 4,945,050, 5,036,006 and 5,100,792, all to Sanford et al., which are hereby incorporated by reference. Generally, this procedure involves propelling inert or biologically active particles at the cells under conditions effective to penetrate the outer surface of the cell and to be incorporated within the interior thereof. When inert particles are utilized, the vector can be introduced into the cell by coating the particles with the vector. Alternatively, the target cell can be surrounded by the vector so that the vector is carried into the cell by the wake of the particle. Biologically active particles (e.g., dried yeast cells, dried bacterium or a bacteriophage, each containing DNA material sought to be introduced) can also be propelled into plant cells. It is not intended, however, that the present invention be limited by the choice of vector or host cell. It should of course be understood that not all vectors and expression control sequences will function equally well to express the DNA sequences of this invention. Neither will all hosts function equally well with the same vector expression system. However, one of skill in the art may make a selection among vectors, expression control sequences, and hosts without undue experimentation and without departing from the scope of this invention.

[0109] An isolated DNA construct selected in accordance with the present invention may be utilized in an expression vector to transform a wide variety of plants, including monocots and dicots. The invention finds advantageous use, for example, in transforming the following plants: rice, wheat, barley, rye, corn, potato, carrot, sweet potato, bean, pea, chicory, lettuce, cabbage, cauliflower, broccoli, turnip, radish, spinach, asparagus, onion, garlic, eggplant, pepper, celery, squash, pumpkin, zucchini, cucumber, apple, pear, quince, melon, plum, cherry, peach, nectarine, apricot, strawberry, grape, raspberry, blackberry, pineapple, avocado, papaya, mango, banana, soybean, tobacco, tomato, sorghum and sugarcane. Additional literature describing plant and/or microorganism transformation includes the following, each of which is incorporated herein by reference in its entirety: Zhijian Li et al. “A Sulfonylurea Herbicide Resistance Gene from Arabidopsis thaliana as a New Selectable Marker for Production of Fertile Transgenic Rice Plants” Plant Physiol. 100, 662-668 (1992); Parsons et al. (1997) Proc. Natl. Acad. Sci. USA 84:4161-4165; Daboussi et al. (1989) Curr. Genet. 15:453-456; Leung et al. (1990) Curr. Genet. 17:409-411; Koetter et al., “Isolation and characterization of the Pichia stipitis xylitol gehydrogenase gene, XYL2, and construction of a xylose-utilizing Saccharomyces cerevisiae transformant,” Curr. Genet., 18:493-500 (1990); Strasser et al., “Cloning of yeast xylose reductase and xylitol dehydrogenase genes and their use,” German patent application (1990); Hallborn et al., “Xylitol production by recombinant Saccharomyces cerevisiae,” Bio./Technol., 9:1090 (1991); Becker and Guarente, “High efficiency transformation of yeast by electroporation,” Methods in Enzymol. 194:182-186 (1991); Ammerer, “Expression of genes in yeast using the ADC1 promoter,” Methods in Enzymol. 101:192-201 (1983); Sarthy et al., “Expression of the E. coli xylose isomerase gene in S. cerevisiae,” Appl. Environ. Microb., 53: 1996′-2000 (1987); U.S. Pat. Nos. 4,945,050, 5,141,131, 5,177,010, 5,104,310, 5,149,645, 5,469,976, 5,464,763, 4,940,838, 4,693,976, 5,591,616, 5,231,019, 5,463,174, 4,762,785, 5,004,863, 5,159,135, 5,302,523, 5,464,765, 5,472,869, 5,384,253; European Patent Application Nos. 0131624B 1, 120516, 159418B1, 176112, 116718, 290799, 320500, 604662, 627752, 0267159, 0292435; WO 87/06614; WO 92/09696; and WO 93/21 335.

[0110] Those skilled in the art will recognize the commercial and agricultural advantages inherent in plants transformed to express feedback insensitive TD. Such plants have the improved ability to synthesize lie and, therefore, are expected to be more valuable nutritionally, compared to a corresponding non-transformed plant. Further, certain intermediates of the Ile biosynthetic pathway have significant commercial value, and production of these intermediates is advantageously increased in a transformant in accordance with the invention. For example, 2-oxobutyrate, the reaction product of the reaction catalyzed by TD, is known to be a precursor for the production of polyhydroxybutyrate in plants that have been genetically engineered using techniques known in the art to include bacterial genes necessary to produce polyhydroxybutyrate. Polyhydroxybutyrate is a desired biopolymer in the plastic industry because it may be biologically degraded. Because plants and microorganisms transformed in accordance with the invention feature increased production of 2-oxobutyrate, such plants and/or microorganisms may be advantageously utilized by plastic manufacturers in this manner. For example, plants that overproduce 2-oxobutyrate would be ideal for metabolic engineering by bacterial genes for polyhydroxybutyrate production because the overproduction of 2-oxobutyrate would provide plenty of substrate for both the natural Ile biosynthetic pathway and the engineered polyhydroxybutyrate pathway.

[0111] Perhaps the most significant advantage of the present invention is that an inventive nucleotide sequence may be used in an expression vector as a selectable marker. In this aspect of the invention, an inventive nucleotide sequence is incorporated into a vector such that it is expressed in a cell transformed thereby, along with a second pre-selected nucleotide sequence (i.e., the primary sequence) which is desired to be incorporated into the genome of the target cell. In this inventive selection protocol, successful transformants will not only express the primary sequence, but will also express a feedback insensitive TD. Thus, once the recombinant DNA is introduced into the plant tissue or microorganism, successful transformants can be screened in accordance with the invention by growing the plant or microorganism in a substrate comprising a toxic Ile analog, such as, for example, OMT (termed “toxic substrate” herein). The Ile structural analog is toxic to wild-type TD, and only the successful transformants, i.e., those expressing feedback insensitive TD, will live, grow and/or proliferate in the toxic substrate.

[0112] In this manner, omr1 is also an excellent biochemical marker to be used in experiments of genetic engineering of bacteria replacing the traditionally used and environmentally-hazardous antibiotic-resistant genes (such as ampicillin- and kanamycin-resistant marker genes), omr1 is very environmentally friendly and poses no risk to human health when included in a transformant, because it does not have an ortholog in humans. Humans do not synthesize isoleucine and may only obtain it by digesting food.

[0113] Based upon the advantageous features of the invention, there is also provided a novel herbicide system. In accordance with this system, agriculturally valuable plant lines comprising an expressible nucleotide sequence encoding an insensitive TD (“transformed plant line”) are grown in a substrate and an Ile structural analog selected in accordance with the invention is contacted with the substrate or with the plants themselves. As a result, only the transformed plants will continue to grow and other plants contacted with the analog will die.

[0114] The invention will be further described with reference to the following specific Examples. It will be understood that these Examples are illustrative and not restrictive in nature. Restriction enzyme digestions, phosphorylations, ligations and bacterial transformations were done as described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition (1989) Cold Spring Harbor Laboratory Press. Plant transformations were done according to Bent et al. “RPS2 of Arabidopsis thaliana: A leucine-rich repeat class of plant disease resistance genes.” Science 265:1856-1860 (1994). Each reference is incorporated herein by reference in its entirety.

EXAMPLE ONE

[0115] As reported in Mourad G, King J (1995) L-O-methylthreonin-resistant mutant of Arabidopsis defective in isoleucine feedback regulation. Plant Physiol 107:43-52, the mutated line GM11b of Arabidopsis thaliana was obtained, using EMS-mutagenesis, by selection in the presence of the toxic Ile structural analog, L-O-methylthreonine (OMT). The basis of mutant selection was that OMT is incorporated into cellular proteins in place of Ile, causing loss of protein function and, thus, cell death. GM11b was rescued because of a dominant mutation in the single gene omr1 which encodes TD. The mutation in the omr1 gene causes TD from GM11b to be insensitive to feedback control by Ile. TD activity in extracts from GM11b plants was about 50-fold more resistant to feedback inhibition by Ile than TD in extracts from wild type plants. The loss of Ile feedback sensitivity in GM11b led to a 20-fold overproduction of free Ile when compared to the wild type. This overproduction of Ile in GM11b had no effect on plant growth or reproduction.

EXAMPLE TWO Cloning, Sequencing and Testing omr1 as a Selectable Marker in Genetic Engineering Experiments

[0116] 1. The construction of a cDNA Library from GM11b (omr1/omr1):

[0117] Total RNA was extracted from 16-day-old GM11b (omr1/omr1) plants that were germinated in a minimal agar medium supplemented with 0.2 mM MTR. Poly(A) RNA (mRNA) was extracted from the total RNA and complementary DNA (cDNA) was synthesized using reverse transcriptase. The cDNA library was synthesized using the ZAP-cDNA synthesis kit of Stratagene. To prime the cDNA synthesis, a 50-base oligonucleotide linker primer containing an Xho I site and an 18-base poly(dT) was used. A 13-mer oligonucleotide adaptor containing an Eco RI cohesive end was ligated to the double stranded cDNA molecules at the 5′ end. This allowed unidirectional cloning of the cDNA molecules, in the sense orientation, into the Eco RI and Xho I sites of the Uni-ZAP XR vector of Stratagene. The recombinant X phage library was amplified using the XL1-Blue MRF′ E. coli host cells yielding a titer 6.8×10⁹ pfu/ml. The average size insert was approximately 1.4 kb. This was calculated from PCR analysis of 20 random, clear plaques isolated from the amplified library. The Uni-ZAP XR vector contains the pBluescript SK(−) plasmid containing the N-terminus of the lacZ gene. To excise the pBluescript phagemid, containing the cloned cDNA insert, the ExAssist/SOLR system provided by Stratagene was used. This allowed the rescue of the cDNA inserts from the positive X clones in pBluescritpt SK plasmids in a single step.

[0118] 2. The isolation of a Small TD-DNA Fragment to Use as a Homologous Probe:

[0119] To isolate the omr1 gene encoding TD from the cDNA library of the line GM11b, a homologous oligonucleotide, isolated from Arabidopsis DNA, was used as a probe against the cDNA library. Taking into consideration that TD is conserved in a variety of organisms, degenerate primers were designed from conserved amino acid regions of TD. Such conserved regions were identified by aligning the amino acid sequence of TD from chickpea and tomato. FIG. 2 shows the location of the conserved amino sequences in tomato and chickpea and also the location of the degenerate oligonucleotide primers TD205 and TD206 that were designed to isolate a TD-DNA fragment from Arabidopsis. FIG. 4 shows the structure and degree of degeneracy of the PCR oligonucleotide primers, TD205 (the 5′ end primer) and TD206 (the 3′ end primer). Both primers TD 205 and TD 206 were designed to accommodate the Arabidopsis codon usage bias. Primer TD 205 had 384-fold degeneracy and was a 28-mer anchored with an Eco RI site starting 2 bases downstream from the first nucleotide at the 5′ end of the primer. TD 206 had 324-fold degeneracy and was a 28-mer anchored with a Hind III site starting 2 bases downstream from the first nucleotide at the 5′ end of the primer.

[0120] Genomic DNA was isolated from GM11b and used as a template in a PCR amplification with the primers TD205 and TD 206. A 438 bp fragment was amplified. The fragment was cloned into the Eco RI-Hind III sites of the plasmid pGEM3Zf(+). The fragment was sequenced to completion using the dideoxy chain termination method and the sequenase kit of USB. The fragment showed a putative 280 bp intron. The remaining 158 bp of the PCR-fragment had 60.1% identical nucleotide sequence with the chickpea TD gene. To eliminate the putative intron sequences, a second pair of primers TD 211 and TD212 were designed and used in a PCR reaction with the 438 bp fragment as a template. A DNA fragment of about 100 bp length, containing exon sequences, was amplified and purified. This was the homologous probe used for screening the cDNA library constructed from GM11b.

[0121] 3. Screening the cDNA Library of GM11b:

[0122] The 100 bp PCR-fragment was labeled with [α-³²P]dCTP (3000 Ci/mmol) using random priming (prime-a gene labeling kit of Promega) and used as a probe to screen plaque lifts (two replicas per plate) of the plated GM11b cDNA library. Hybridization was done at 42° C. in formamide for 2 days. The nylon membranes containing the plaque lifts were washed 3×at room temperature (25° C.) in 7×SSPE and 0.5%SDS for 5 minutes. The nylon membranes were then put on X-ray film and exposed for 1 day. Two plaques hybridized and showed signal on the X-ray films of the two replicas taken from the same plate. At the site of positive hybridization, plugs were cut out of the agar plate and put in 1 ml of SM buffer with 20 μL chloroform. A secondary, tertiary and quaternary screening was performed until about 90% of the plaques on the plate showed a strong signal on the X-ray film of both replicas of the same plate. A well isolated plaque representing each clone was cut out from the plate and put in SM buffer. The phage eluate was infected with the ExAssist helper phage to excise the pBluescript SK plasmid containing the cDNA insert and the resulting recombinant bacteria was plated on media with ampicillin (60 μg/ml). A few bacterial colonies were selected, plasmid DNA was prepared then digested with Eco RI and Xho I to release the inserts. A Southern blot was prepared from the plasmid digests and probed with the ³²P-labelled 100 bp TD fragment. All the clones, descendants from the two phage clones, showed very strong signal. This was a strong indication that the isolated clones contained the TD from the line GM11b. One clone was named TD23 and was selected for DNA sequencing. The size of the cDNA insert in clone TD23 was 2229 nucleotides.

[0123] 4. Sequencing of the 2229 bp Fragment of the Clone TD23:

[0124] Sequencing of the cDNA insert of clone TD23 was performed by the dideoxy chain termination method using the sequenase kit of USB. To start the sequencing project, an oligonucleotide primer complementary to the T3 promoter of pBluescript SK was synthesized and used to obtain the sequence of the first few nucleotides of the insert. This sequence, 30 nucleotides, included the multiple cloning site downstream of the T3 promoter. The start of the cDNA sequence was immediately following the Eco RI site which starts at position 31. DNA sequencing was also performed on the opposite strand starting from the 3′ end and using the T7 promoter of the pBluescript SK. Both strands of the TD 23 insert were sequenced to completion using a set of oligonucleotide primers designed from the DNA revealed after each sequencing reaction. A total of 19 oligonucleotide primers were synthesized and used in sequencing the cDNA insert.

[0125] The total length of the sequenced fragment was 2277 nucleotides of which 2229 were the cDNA insert. Of the remaining 48 nucleotides, 2277-2229, 31 nucleotides were the multiple cloning site between the T3 promoter and the Eco RI site at the 5′ end of the insert and 17 nucleotides were multiple cloning site between the T7 promoter and Xho I site at the 3′ end of the insert (FIG. 4). FIG. 5 shows the nucleotide sequence and the predicted amino acid sequence of clone 23 as isolated from the cDNA library constructed from line GM11b of Arabidopsis (omr1/omr1). The TD insert in clone 23 is in pBluescript vector between the Eco RI and Xho I sites. An open reading frame (top reading frame) was observed which showed an ATG codon at nucleotide 166 and a termination codon at nucleotide 1801. The total cDNA insert in clone 23 is 1758 nucleotides (including the stop codon) encoding a polypeptide of 585 amino acids. FIG. 4 shows the DNA sequence of clone 23 and FIG. 5 shows the DNA sequence and the open reading frame with the predicted amino acid sequence encoded by the cDNA insert. The predicted amino acid sequence encoded by the TD 23 cDNA gene shared greater than 50% identity with the amino acid sequence of TD of potato and tomato respectively. This was strong evidence that the cDNA insert of the clone TD23 is indeed the gene encoding threonine dehydratase/deaminase, omr1, of the L-O-methylthreonine-resistant line GM1 lb of Arabidopsis thaliana.

[0126] 5. Test of Functionality of the cDNA Insert (omr1) Encoding TD of Arabidopsis:

[0127] To test that the cloned cDNA insert of the clone TD 23 is indeed encoding a functional threonine dehydratase/deaminase, a complementation test was performed. The E. coli strain TGXA is an auxotroph with a deletion in the ilvA gene encoding threonine dehydratase/dearminase. Fisher K E, Eisenstein e (1993) An efficient approach to identify ilva mutations reveals an amino-terminal catalytic domain in biosynthetic threonine deaminase from Escherichia coli. J Bacteriol 175:6605-6613. This strain cannot grow on a minimal medium without supplementation with Ile. This strain was a generous gift from Drs. Kathryn E. Fisher and Edward Eisenstein, University of Maryland Baltimore County, Maryland.

[0128] First complementation experiments were done to test the ability of omr1 to revert the bacterial Ile auxotroph TGXA to prototrophy. This was done by transforming TGXA with pGM-td23, containing the cDNA insert omr1 in pBluescript SK under the control of the T3 promoter. In addition, the cDNA insert containing omr1 was subcloned in two different prokaryotic expression vectors. An Xba I-Xho I fragment, containing the cDNA sequence of omr1, was excised from pGM-td23 and cloned into Xba 1-Sal I linearized prokaryotic expression vectors pTrc99A and pUCK2. In pTrc99A, omr1 was cloned in front of the lacZ IPTG-inducible promoter while in pUCK2, omr1 was cloned in front of a constitutive promoter. Xho I and Sal I cohesive termini are compatible and therefore allowed the ligation of the inserts into the expression vectors. The recombinant vectors pTrc-td23, pUCK-td23 or pBluescript-td23 all containing full length omr1 were transformed into the strain TGXA and plated on minimal media without supplementation. All of the three constructs were able to revert Ile auxotrophy of the host TGXA to prototrophy. These experiments confirmed that omr1 encoding Arabidopsis thaliana (line GM11b) TD is functional and able to unblock the Ile biosynthetic pathway of the E. coli strain TGXA.

[0129] In the second complementation experiment, the E. coli prototroph host DH5α was transformed with pTrc-td23 or pUCK-td23 and plated on minimal medium supplemented with varying concentrations of the toxic analog L-O-methylthreonine. Both of the constructs were able to confer upon DH5α resistance to 30 μM L-O-methylthreonine. No bacterial colonies grew on plates containing untransformed DH5a. This result provided strong evidence that the mutated omr1 gene of the line GM11b of Arabidopsis is able to confer resistance to L-O-methylthreonine present in the growth medium. Therefore omr1 provides a new environmentally friendly selectable marker for genetic transformation of bacteria.

[0130] 6. Construction of the pCM35S-omr1 Expression Vector for Plant Transformation:

[0131] The strategy for cloning the omr1. allele into a plant expression vector was as follows:

[0132] A. The coding region of omr1 allele was excised from pGM-td23 as an Xba I-Kpn I fragment.

[0133] B. The 500 bp CaMV 35S promoter was cleaved out of the vector pBI121.1 (Jefferson et al., 1987) with Hind III and Bam HI. The pBIN19 vector was linearized by Hind III and Bam HI then ligated to the CaMV 35S promoter so as to place the promoter into the multiple cloning site in the correct orientation. This vector was named pCM35S.

[0134] C. The plasmid pCM35S was digested with Xba I-Kpn I and the omr1 fragment isolated in step A was cloned into the Xba I-Kpn I sites placing the omr1 coding sequence in front of the CaMV 35S promoter and creating a plasmid with the kanamycin resistance gene (NOS:NPT11:NOS) close to the right border RB of the T-DNA region of the Ti plasmid and 35S:omr1 downstream and close to the left border LB of the T-DNA region of the Ti plasmid. This plasmid was named pCM35S-omr1-nos (ca. 13 kb).

[0135] D. The NOS terminator of pBIN19 was PCR-amplified using a pair of oligonucleotide primers, the 5′ primer was anchored with an Xba I site and the 3′ primer was anchored with a Sal I site. PCR amplification yielded a 300 bp NOS terminator fragment.

[0136] E. To clone a NOS terminator to the 3′ end of the omr1 gene, the recombinant plasmid pCM35S-omr1-nos was digested with Nhe Iand Xho 1. This yielded three fragments:

[0137] (i) a 5 kb Nhe I-Nhe I fragment containing part of the NOS promoter of the NPTII gene, the 35S promoter and the full length omr1 cDNA except 200 bp of non-translated sequences at the 3′ end which include the poly A tail.

[0138] (ii) a 200 bp Nhe I-Xho I fragment containing the 200 bp fragment mentioned in (i) and that contained the poly A tail and non-translated sequences at the 3′ end of omr1.

[0139] (iii) an 8 kb Nhe I-Xho I fragment containing the 5′ end NOS promoter of the NPTII gene and the remaining sequences outside LB and RB of the pCM35S-omr1-nos.

[0140] F. To clone the NOS terminator immediately downstream from the omr1 gene in pCM35S-omr1-nos, a triple ligation was performed including the 5 kb Nhe I-Nhe I fragment containing part of the NOS promoter of the NPTII gene mentioned above in E(i), the 300 bp Xba I-Sal I NOS terminator fragment mentioned in C, and the 8 kb Nhe I-Xho I fragment containing the 5′ end NOS promoter of the NPTII gene and the remaining sequences outside LB and RB of the pCM35S-omr1-nos. The result of this triple cloning was the ligation of the 5 kb fragment at one Nhe I end (the NOS promoter end) to the Nhe I site of the 8 kb fragment (Nhe I/Nhe I) and the other Nhe I end (at the 3′ end of the omr1 coding sequence) of the 5 kb fragment was ligated to the Xba I (isoschizomer) of the 300 bp NOS terminator fragment. The Sal I end of the 300 bp NOS terminator was ligated to the Xho I (isoschizomer) end of the 8 kb fragment. This generated the recombinant plasmid pCM35S-omr1 containing the omr1 gene driven by the CaMV 35S promoter and terminated by the NOS terminator and the kanamycin resistance gene (NOS promoter:NPTII:NOS:terminator) between the LB and RB (FIG. 16). To confirm the cloning of the three fragments in the proper orientation, a diagnostic digestion with Xba I & Kpn I produced a 2.3-2.4 kb fragment. The plasmid pCM35S-omr1 therefore contained two constructs that could be expressed in plants, the CaMV35S:omr1:NOS terminator expressing L-O-methylthreonine-resistance and the NOS promoter:NPT11:NOS terminator expressing kanamycin-resistance.

[0141] 7. Plant Transformation Using pCM35S-omr1:

[0142] Using the vacuum infiltration method of Bent et al. (1994), L-O-methylthreonine-sensitive Arabidopsis thaliana Columbia wild type were transformed with pCM35S-omr1. Ten pots, each with 3-4 plants, were transformed and T1 seeds were harvested from the T_(o) transformed plants of each pot separately. The T1 seeds from each pot were screened for expression of L-O-methylthreonine resistance by germinating in agar medium supplemented with 0.2 mM L-O-methylthreonine, a concentration previously determined and known to completely inhibit the growth of wild type seedlings beyond the cotyledonous stage (Mourad and King, 1995). Half of the T1 seeds from each of the ten pots were screened for L-O-methylthreonine resistance and 5 independent transformants were able to germinate and continue to grow healthy roots and shoots among thousands of seedlings that were completely bleached immediately after the emergence of the cotyledons. In a crowded plate, it is possible to identify the transformants by looking at the bottom of the plate, the transformants show root growth while the nontransformants will have none. After three weeks of growth in the 0.2 mM L-O-methylthreonine agar medium, each of the 5 positive transformants was transferred to soil, kept separately and allowed to self-fertilize to produce the T2 seed.

[0143] 8. Genetic Characterization of the omr1 Transformants:

[0144] The T2 seed was harvested from each of the 5 positive T1 transformants and 50 T2 seeds/transformant were planted in a separate petri plate containing 0.2 mM L-O-methylthreonine agar medium. In each of the 5 petri plates, the majority (75% or more) of the T2 seedlings were resistant to L-O-methylthreonine indicating that a single copy of the transgene omr1 had been inserted in the parent T1 transgenic plant. FIG. 6b shows that 585 amino acid residues of the total 592 residues representing the full length mutant TD were expressed in the transgenic plants. This slightly truncated precursor mutant TD was able to translocate to the chloroplast and confer upon transgenic plants resistance to OMT.

[0145] 9. Molecular Characterization of the omr1 Transformants:

[0146] Two to three leaves of each of the five T1 transformants was excised from the plants at the rosette stage and total DNA was extracted according to a modification of the procedure of Konieczny and Ausubel (1993). A PCR approach was used to confirm the presence of the introduced transgene omr1. For that, a pair of oligonucleotide primers were synthesized such that one primer is complementary to the start of the omr1 and the other primer was complementary to the end of the NOS terminator. The PCR reaction using DNA extracted from each of the five T1 transformants was PCR amplified and each produced a 2.5 kb fragment confirming the presence of the transgene omr1 followed by the NOS terminator in each of the transformants. The native wild type allele OMR1 did not PCR amplify because it is not followed by the NOS terminator and therefore no PCR reaction could take place. DNA extracted from untransformed Arabidopsis plants failed to amplify using such primers.

EXAMPLE THREE The Molecular Basis of L-O-Methylthreonine Resistance Encoded by the omr1 Allele of Line GM11b of Arabidopsis thaliana

[0147] 1. Isolation of the Wild Type OMR1 Allele:

[0148] An Arabidopsis thaliana Columbia wild type cDNA library constructed from 3-day-old seedlings in Stratagene's λ ZAP II vector was screened with a ³²P-labeled 1080 base pair DNA fragment PCR-amplified from the cDNA sequence of omr1 (described above) as a probe. The screening yielded a positive clone TD54 which was purified and was proven to be the wild type allele OMR1 by PCR and Southern analysis.

[0149] 2. Sequencing of the OMR1 Wild Type Allele:

[0150] The recombinant plasmid containing the wild type allele OMRI was named pGM-td54 and the OMR1 allele was manually sequenced using the sequenase kit of USB and the same set of oligonucleotide primers that were previously used in sequencing the omr1 allele. The DNA sequence of the wild type OMR1 was similar to that of omr1 except for two different base substitutions predicting two amino acid substitutions in the mutated TD encoded by omr1. In an attempt to clone the 5′ upstream sequences from the ATG start codon of clone 23 (FIG. 5) and using a PCR approach, a new ATG codon was detected at 141 nucleotides upstream from the ATG codon reported in clone 23. This was confirmed in both the wild type allele OMR1 and the mutated allele omr1. Therefore the full length cDNA of the omr1 locus was found to be 1779 nucleotides (FIG. 7) encoding a TD protein of 592 amino acids (FIGS. 8 and 9). The omr1 insert as shown in FIG. 6b (SEQ ID NO:3) was not only strongly expressed in the first transgenic plants (T1) but was also inherited and strongly expressed in their progeny (T2 plants). As expected, the full length cDNA of the OMR1 allele of the omr1 locus was 1779 nucleotides (FIG. 10) encoding a wild type TD of 592 amino acids (FIGS. 11 and 12).

[0151] Amino acid alignment of wild type threonine dehydratase/deaminase of Arabidopsis thaliana with that of chickpea (John et al., 1995), tomato (Samach et al., 1991), potato (Hildmann T, Ebneth M, Pena-Cortes H, Sanchez-Serrano J J, Willmitzer L, Prat S (1992) General roles of abscisic and jasmonic acids in gene activation as a result of mechanical wounding. Plant Cell 4:1157-1170.), yeast I (Kielland-Brandt M C, Holmberg S, Petersen J G L, Nilsson-Tillgren T (1984) Nucleotide sequence of the gene for threonine deaminase (ilvl) of Saccharomyces cerevisiae. Carlsberg Res Commun 49:567-575.), yeast 2 (Bornaes C, Petersen J G, Holmberg S (1992) Serine and threonine catabolosm in Saccharomyces cerevisiae: the CHA 1 polypeptide is homologous with other serine and threonine dehydratases. Genetics 131:531-539.), E. coli biosynthetic (Wek R C, Hatfield G C (1986) Nucleotide sequence and in vivo expression of ilvY and ilvC genes in Escherichia coli K12. Transcription from divergent overlapping promoters. J Biol Chem 261:2441-2450.), E. coli catabolic (Datta P, Goss T J, Omnaas J R, Patil R V (1987) Covalent structure of biodegradative threonine dehydratase of Escherichia coli: homology with other dehydratases. Proc Natl Acad Sci USA 84:393-397.), and Salmonella typhimurium (Taillon B E, Little R, Lawther R P (1988) Analysis of the functional domains of biosynthetic threonine deaminase by comparison of the amino acid sequences of three wild type alleles to the amino acid of biodegradative threonine deaminase. Gene 62:245-252.) is set forth in FIG. 13. The Megalign program of the Lasergene software was used, DNASTAR Inc., Madison, Wis. The degree of similarity between amino acid residues of Arabidopsis threonine dehydratase/deaminase and those of other organisms was calculated by the Lipman-Pearson protein alignment method using the Lasergene software and was found to be 46.2% with chickpea, 52.7% with tomato, 55.0% with potato (partial), 45.0% with yeast 1, 24.7% yeast 2, 43.4% with E. coli (biosynthetic), 39.3% with E. coli (catabolic) and 43.3% with Salmonella.

[0152] 3. Comparing DNA Sequences of omr1 and OMR1 Revealed the Point Mutations Involved:

[0153] With reference to the nucleotide residue numbering in SEQ ID NO:1 and SEQ ID NO:2, the first base substitution occurred at nucleotide 1519 where C (cytosine) in the wild type allele OMR1 was substituted by T (thymine) in the mutated allele omr1 (FIGS. 14 & 15). This base substitution predicted an amino acid substitution at amino acid residue 452 at the polypeptide level where the arginine residue in the wild type TD encoded by OMR1 was substituted by a cysteine residue in the mutated isoleucine-insensitive TD encoded by omr1 (FIG. 15). This point mutation resides in a conserved regulatory region of amino acids designated R4 (regulatory) by Taillon et al. (1988) where the mutated amino acid is normally an arginine residue in the TD of Arabidopsis, yeast 1, E. coli (biosynthetic) and Salmonella and a lysine residue in the TD of chickpea, tomato, and potato (partial) (FIG. 16). The second base substitution occurred at nucleotide 1655 where G (guanine) in the wild type allele OMR1 was substituted by A (adenine) in the mutated allele omr1 (FIGS. 17 & 18). This base substitution predicted an amino acid substitution at residue 597 at the polypeptide level where the arginine residue in the wild type TD encoded by OMR1 was substituted by a histidine residue in the mutated isoleucine-insensitive TD encoded by omr1 (FIG. 18). This point mutation resides in a conserved regulatory region of amino acids designated R6 (regulatory) by Taillon et al. (1988) where the mutated amino acid is normally an arginine residue in TD of Arabidopsis, chickpea, tomato, potato (partial), yeast 1, E. coli (biosynthetic) and Salmonella (FIG. 19).

1 69 1 1779 DNA Arabidopsis thaliana CDS (1)..(1779) 1 atg aat tcc gtt cag ctt ccg acg gcg caa tcc tct ctc cgt agc cac 48 Met Asn Ser Val Gln Leu Pro Thr Ala Gln Ser Ser Leu Arg Ser His 1 5 10 15 att cac cgt cca tca aaa cca gtg gtc gga ttc act cac ttc tcc tcc 96 Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser Ser 20 25 30 cgt tct cgg atc gca gtg gcg gtt ctg tcc cga gat gaa aca tct atg 144 Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser Met 35 40 45 act cca ccg cct cca aag ctt cct tta cca cgt ctt aag gtc tct ccg 192 Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser Pro 50 55 60 aat tcg ttg caa tac cct gcc ggt tac ctc ggt gct gta cca gaa cgt 240 Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu Arg 65 70 75 80 acg aac gag gct gag aac gga agc atc gcg gaa gct atg gag tat ttg 288 Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr Leu 85 90 95 acg aat ata ctg tcc act aag gtt tac gac atc gcc att gag tca cca 336 Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser Pro 100 105 110 ctc caa ttg gct aag aag cta tct aag aga tta ggt gtt cgt atg tat 384 Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met Tyr 115 120 125 ctt aaa aga gaa gac ttg caa cct gta ttc tcg ttt aag ctt cgt gga 432 Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg Gly 130 135 140 gct tac aat atg atg gtg aaa ctt cca gca gat caa ttg gca aaa gga 480 Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys Gly 145 150 155 160 gtt atc tgc tct tca gct gga aac cat gct caa gga gtt gct tta tct 528 Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu Ser 165 170 175 gct agt aaa ctc ggc tgc act gct gtg att gtt atg cct gtt acg act 576 Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr Thr 180 185 190 cct gag ata aag tgg caa gct gta gag aat ttg ggt gca acg gtt gtt 624 Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val Val 195 200 205 ctt ttc gga gat tcg tat gat caa gca caa gca cat gct aag ata cga 672 Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile Arg 210 215 220 gct gaa gaa gag ggt ctg acg ttt ata cct cct ttt gat cac cct gat 720 Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro Asp 225 230 235 240 gtt att gct gga caa ggg act gtt ggg atg gag atc act cgt cag gct 768 Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln Ala 245 250 255 aag ggt cca ttg cat gct ata ttt gtg cca gtt ggt ggt ggt ggt tta 816 Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly Leu 260 265 270 ata gct ggt att gct gct tat gtg aag agg gtt tct ccc gag gtg aag 864 Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val Lys 275 280 285 atc att ggt gta gaa cca gct gac gca aat gca atg gct ttg tcg ctg 912 Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser Leu 290 295 300 cat cac ggt gag agg gtg ata ttg gac cag gtt ggg gga ttt gca gat 960 His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala Asp 305 310 315 320 ggt gta gca gtt aaa gaa gtt ggt gaa gag act ttt cgt ata agc aga 1008 Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser Arg 325 330 335 aat cta atg gat ggt gtt gtt ctt gtc act cgt gat gct att tgt gca 1056 Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys Ala 340 345 350 tca ata aag gat atg ttt gag gag aaa cgg aac ata ttg gaa cca gca 1104 Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala 355 360 365 ggg gct ctt gca ctc gct gga gct gag gca tac tgt aaa tat tat ggc 1152 Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly 370 375 380 cta aag gac gtg aat gtc gta gcc ata acc agt ggc gct aac atg aac 1200 Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn 385 390 395 400 ttt gac aag cta agg att gtg aca gaa ctc gcc aat gtc ggt agg caa 1248 Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln 405 410 415 cag gaa gct gtt ctt gct act ctc atg ccg gaa aaa cct gga agc ttt 1296 Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe 420 425 430 aag caa ttt tgt gag ctg gtt gga cca atg aac ata agc gag ttc aaa 1344 Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys 435 440 445 tat aga tgt agc tcg gaa aag gag gct gtt gta cta tac agt gtc gga 1392 Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly 450 455 460 gtt cac aca gct gga gag ctc aaa gca cta cag aag aga atg gaa tct 1440 Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser 465 470 475 480 tct caa ctc aaa act gtc aat ctc act acc agt gac tta gtg aaa gat 1488 Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp 485 490 495 cac ctg cgt tac ttg atg gga gga aga tct act gtt gga gac gag gtt 1536 His Leu Arg Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 500 505 510 cta tgc cga ttc acc ttt ccc gag aga cct ggt gct cta atg aac ttc 1584 Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe 515 520 525 ttg gac tct ttc agt cca cgg tgg aac atc acc ctt ttc cat tac cgt 1632 Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr Arg 530 535 540 gga cag ggt gag acg ggc gcg aat gtg ctg gtc ggg atc caa gtc ccc 1680 Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro 545 550 555 560 gag caa gaa atg gag gaa ttt aaa aac cga gct aaa gct ctt gga tac 1728 Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr 565 570 575 gac tac ttc tta gta agt gat gac gac tat ttt aag ctt ctg atg cac 1776 Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 580 585 590 tga 1779 2 592 PRT Arabidopsis thaliana 2 Met Asn Ser Val Gln Leu Pro Thr Ala Gln Ser Ser Leu Arg Ser His 1 5 10 15 Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser Ser 20 25 30 Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser Met 35 40 45 Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser Pro 50 55 60 Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu Arg 65 70 75 80 Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr Leu 85 90 95 Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser Pro 100 105 110 Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met Tyr 115 120 125 Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg Gly 130 135 140 Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys Gly 145 150 155 160 Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu Ser 165 170 175 Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr Thr 180 185 190 Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val Val 195 200 205 Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile Arg 210 215 220 Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro Asp 225 230 235 240 Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln Ala 245 250 255 Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly Leu 260 265 270 Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val Lys 275 280 285 Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser Leu 290 295 300 His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala Asp 305 310 315 320 Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser Arg 325 330 335 Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys Ala 340 345 350 Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala 355 360 365 Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly 370 375 380 Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn 385 390 395 400 Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln 405 410 415 Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe 420 425 430 Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys 435 440 445 Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly 450 455 460 Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser 465 470 475 480 Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp 485 490 495 His Leu Arg Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 500 505 510 Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe 515 520 525 Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr Arg 530 535 540 Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro 545 550 555 560 Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr 565 570 575 Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 580 585 590 3 1779 DNA Arabidopsis thaliana CDS (1)..(1779) 3 atg aat tcc gtt cag ctt ccg acg gcg caa tcc tct ctc cgt agc cac 48 Met Asn Ser Val Gln Leu Pro Thr Ala Gln Ser Ser Leu Arg Ser His 1 5 10 15 att cac cgt cca tca aaa cca gtg gtc gga ttc act cac ttc tcc tcc 96 Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser Ser 20 25 30 cgt tct cgg atc gca gtg gcg gtt ctg tcc cga gat gaa aca tct atg 144 Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser Met 35 40 45 act cca ccg cct cca aag ctt cct tta cca cgt ctt aag gtc tct ccg 192 Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser Pro 50 55 60 aat tcg ttg caa tac cct gcc ggt tac ctc ggt gct gta cca gaa cgt 240 Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu Arg 65 70 75 80 acg aac gag gct gag aac gga agc atc gcg gaa gct atg gag tat ttg 288 Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr Leu 85 90 95 acg aat ata ctg tcc act aag gtt tac gac atc gcc att gag tca cca 336 Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser Pro 100 105 110 ctc caa ttg gct aag aag cta tct aag aga tta ggt gtt cgt atg tat 384 Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met Tyr 115 120 125 ctt aaa aga gaa gac ttg caa cct gta ttc tcg ttt aag ctt cgt gga 432 Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg Gly 130 135 140 gct tac aat atg atg gtg aaa ctt cca gca gat caa ttg gca aaa gga 480 Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys Gly 145 150 155 160 gtt atc tgc tct tca gct gga aac cat gct caa gga gtt gct tta tct 528 Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu Ser 165 170 175 gct agt aaa ctc ggc tgc act gct gtg att gtt atg cct gtt acg act 576 Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr Thr 180 185 190 cct gag ata aag tgg caa gct gta gag aat ttg ggt gca acg gtt gtt 624 Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val Val 195 200 205 ctt ttc gga gat tcg tat gat caa gca caa gca cat gct aag ata cga 672 Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile Arg 210 215 220 gct gaa gaa gag ggt ctg acg ttt ata cct cct ttt gat cac cct gat 720 Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro Asp 225 230 235 240 gtt att gct gga caa ggg act gtt ggg atg gag atc act cgt cag gct 768 Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln Ala 245 250 255 aag ggt cca ttg cat gct ata ttt gtg cca gtt ggt ggt ggt ggt tta 816 Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly Leu 260 265 270 ata gct ggt att gct gct tat gtg aag agg gtt tct ccc gag gtg aag 864 Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val Lys 275 280 285 atc att ggt gta gaa cca gct gac gca aat gca atg gct ttg tcg ctg 912 Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser Leu 290 295 300 cat cac ggt gag agg gtg ata ttg gac cag gtt ggg gga ttt gca gat 960 His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala Asp 305 310 315 320 ggt gta gca gtt aaa gaa gtt ggt gaa gag act ttt cgt ata agc aga 1008 Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser Arg 325 330 335 aat cta atg gat ggt gtt gtt ctt gtc act cgt gat gct att tgt gca 1056 Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys Ala 340 345 350 tca ata aag gat atg ttt gag gag aaa cgg aac ata ttg gaa cca gca 1104 Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala 355 360 365 ggg gct ctt gca ctc gct gga gct gag gca tac tgt aaa tat tat ggc 1152 Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly 370 375 380 cta aag gac gtg aat gtc gta gcc ata acc agt ggc gct aac atg aac 1200 Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn 385 390 395 400 ttt gac aag cta agg att gtg aca gaa ctc gcc aat gtc ggt agg caa 1248 Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln 405 410 415 cag gaa gct gtt ctt gct act ctc atg ccg gaa aaa cct gga agc ttt 1296 Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe 420 425 430 aag caa ttt tgt gag ctg gtt gga cca atg aac ata agc gag ttc aaa 1344 Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys 435 440 445 tat aga tgt agc tcg gaa aag gag gct gtt gta cta tac agt gtc gga 1392 Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly 450 455 460 gtt cac aca gct gga gag ctc aaa gca cta cag aag aga atg gaa tct 1440 Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser 465 470 475 480 tct caa ctc aaa act gtc aat ctc act acc agt gac tta gtg aaa gat 1488 Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp 485 490 495 cac ctg tgt tac ttg atg gga gga aga tct act gtt gga gac gag gtt 1536 His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 500 505 510 cta tgc cga ttc acc ttt ccc gag aga cct ggt gct cta atg aac ttc 1584 Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe 515 520 525 ttg gac tct ttc agt cca cgg tgg aac atc acc ctt ttc cat tac cat 1632 Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr His 530 535 540 gga cag ggt gag acg ggc gcg aat gtg ctg gtc ggg atc caa gtc ccc 1680 Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro 545 550 555 560 gag caa gaa atg gag gaa ttt aaa aac cga gct aaa gct ctt gga tac 1728 Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr 565 570 575 gac tac ttc tta gta agt gat gac gac tat ttt aag ctt ctg atg cac 1776 Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 580 585 590 tga 1779 4 592 PRT Arabidopsis thaliana 4 Met Asn Ser Val Gln Leu Pro Thr Ala Gln Ser Ser Leu Arg Ser His 1 5 10 15 Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser Ser 20 25 30 Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser Met 35 40 45 Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser Pro 50 55 60 Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu Arg 65 70 75 80 Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr Leu 85 90 95 Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser Pro 100 105 110 Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met Tyr 115 120 125 Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg Gly 130 135 140 Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys Gly 145 150 155 160 Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu Ser 165 170 175 Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr Thr 180 185 190 Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val Val 195 200 205 Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile Arg 210 215 220 Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro Asp 225 230 235 240 Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln Ala 245 250 255 Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly Leu 260 265 270 Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val Lys 275 280 285 Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser Leu 290 295 300 His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala Asp 305 310 315 320 Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser Arg 325 330 335 Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys Ala 340 345 350 Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala 355 360 365 Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly 370 375 380 Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn 385 390 395 400 Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln 405 410 415 Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe 420 425 430 Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys 435 440 445 Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly 450 455 460 Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser 465 470 475 480 Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp 485 490 495 His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 500 505 510 Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe 515 520 525 Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr His 530 535 540 Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro 545 550 555 560 Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr 565 570 575 Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 580 585 590 5 1830 DNA Arabidopsis thaliana CDS (1)..(1830) 5 atg ggc gag ctc ggt acc cgg gga tcc tct aga act agt gga tcc ccc 48 Met Gly Glu Leu Gly Thr Arg Gly Ser Ser Arg Thr Ser Gly Ser Pro 1 5 10 15 ggg ctg cag gaa ttc ggc acg agg acg gcg caa tcc tct ctc cgt agc 96 Gly Leu Gln Glu Phe Gly Thr Arg Thr Ala Gln Ser Ser Leu Arg Ser 20 25 30 cac att cac cgt cca tca aaa cca gtg gtc gga ttc act cac ttc tcc 144 His Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser 35 40 45 tcc cgt tct cgg atc gca gtg gcg gtt ctg tcc cga gat gaa aca tct 192 Ser Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser 50 55 60 atg act cca ccg cct cca aag ctt cct tta cca cgt ctt aag gtc tct 240 Met Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser 65 70 75 80 ccg aat tcg ttg caa tac cct gcc ggt tac ctc ggt gct gta cca gaa 288 Pro Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu 85 90 95 cgt acg aac gag gct gag aac gga agc atc gcg gaa gct atg gag tat 336 Arg Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr 100 105 110 ttg acg aat ata ctg tcc act aag gtt tac gac atc gcc att gag tca 384 Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser 115 120 125 cca ctc caa ttg gct aag aag cta tct aag aga tta ggt gtt cgt atg 432 Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met 130 135 140 tat ctt aaa aga gaa gac ttg caa cct gta ttc tcg ttt aag ctt cgt 480 Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg 145 150 155 160 gga gct tac aat atg atg gtg aaa ctt cca gca gat caa ttg gca aaa 528 Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys 165 170 175 gga gtt atc tgc tct tca gct gga aac cat gct caa gga gtt gct tta 576 Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu 180 185 190 tct gct agt aaa ctc ggc tgc act gct gtg att gtt atg cct gtt acg 624 Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr 195 200 205 act cct gag ata aag tgg caa gct gta gag aat ttg ggt gca acg gtt 672 Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val 210 215 220 gtt ctt ttc gga gat tcg tat gat caa gca caa gca cat gct aag ata 720 Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile 225 230 235 240 cga gct gaa gaa gag ggt ctg acg ttt ata cct cct ttt gat cac cct 768 Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro 245 250 255 gat gtt att gct gga caa ggg act gtt ggg atg gag atc act cgt cag 816 Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln 260 265 270 gct aag ggt cca ttg cat gct ata ttt gtg cca gtt ggt ggt ggt ggt 864 Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly 275 280 285 tta ata gct ggt att gct gct tat gtg aag agg gtt tct ccc gag gtg 912 Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val 290 295 300 aag atc att ggt gta gaa cca gct gac gca aat gca atg gct ttg tcg 960 Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser 305 310 315 320 ctg cat cac ggt gag agg gtg ata ttg gac cag gtt ggg gga ttt gca 1008 Leu His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala 325 330 335 gat ggt gta gca gtt aaa gaa gtt ggt gaa gag act ttt cgt ata agc 1056 Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser 340 345 350 aga aat cta atg gat ggt gtt gtt ctt gtc act cgt gat gct att tgt 1104 Arg Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys 355 360 365 gca tca ata aag gat atg ttt gag gag aaa cgg aac ata ttg gaa cca 1152 Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro 370 375 380 gca ggg gct ctt gca ctc gct gga gct gag gca tac tgt aaa tat tat 1200 Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr 385 390 395 400 ggc cta aag gac gtg aat gtc gta gcc ata acc agt ggc gct aac atg 1248 Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met 405 410 415 aac ttt gac aag cta agg att gtg aca gaa ctc gcc aat gtc ggt agg 1296 Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg 420 425 430 caa cag gaa gct gtt ctt gct act ctc atg ccg gaa aaa cct gga agc 1344 Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser 435 440 445 ttt aag caa ttt tgt gag ctg gtt gga cca atg aac ata agc gag ttc 1392 Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe 450 455 460 aaa tat aga tgt agc tcg gaa aag gag gct gtt gta cta tac agt gtc 1440 Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val 465 470 475 480 gga gtt cac aca gct gga gag ctc aaa gca cta cag aag aga atg gaa 1488 Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu 485 490 495 tct tct caa ctc aaa act gtc aat ctc act acc agt gac tta gtg aaa 1536 Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys 500 505 510 gat cac ctg tgt tac ttg atg gga gga aga tct act gtt gga gac gag 1584 Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu 515 520 525 gtt cta tgc cga ttc acc ttt ccc gag aga cct ggt gct cta atg aac 1632 Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn 530 535 540 ttc ttg gac tct ttc agt cca cgg tgg aac atc acc ctt ttc cat tac 1680 Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr 545 550 555 560 cat gga cag ggt gag acg ggc gcg aat gtg ctg gtc ggg atc caa gtc 1728 His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val 565 570 575 ccc gag caa gaa atg gag gaa ttt aaa aac cga gct aaa gct ctt gga 1776 Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly 580 585 590 tac gac tac ttc tta gta agt gat gac gac tat ttt aag ctt ctg atg 1824 Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met 595 600 605 cac tga 1830 His 6 609 PRT Arabidopsis thaliana 6 Met Gly Glu Leu Gly Thr Arg Gly Ser Ser Arg Thr Ser Gly Ser Pro 1 5 10 15 Gly Leu Gln Glu Phe Gly Thr Arg Thr Ala Gln Ser Ser Leu Arg Ser 20 25 30 His Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser 35 40 45 Ser Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser 50 55 60 Met Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser 65 70 75 80 Pro Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu 85 90 95 Arg Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr 100 105 110 Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser 115 120 125 Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met 130 135 140 Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg 145 150 155 160 Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys 165 170 175 Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu 180 185 190 Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr 195 200 205 Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val 210 215 220 Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile 225 230 235 240 Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro 245 250 255 Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln 260 265 270 Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly 275 280 285 Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val 290 295 300 Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser 305 310 315 320 Leu His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala 325 330 335 Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser 340 345 350 Arg Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys 355 360 365 Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro 370 375 380 Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr 385 390 395 400 Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met 405 410 415 Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg 420 425 430 Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser 435 440 445 Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe 450 455 460 Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val 465 470 475 480 Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu 485 490 495 Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys 500 505 510 Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu 515 520 525 Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn 530 535 540 Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr 545 550 555 560 His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val 565 570 575 Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly 580 585 590 Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met 595 600 605 His 7 1509 DNA Arabidopsis thaliana CDS (1)..(1509) 7 gaa gct atg gag tat ttg acg aat ata ctg tcc act aag gtt tac gac 48 Glu Ala Met Glu Tyr Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp 1 5 10 15 atc gcc att gag tca cca ctc caa ttg gct aag aag cta tct aag aga 96 Ile Ala Ile Glu Ser Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg 20 25 30 tta ggt gtt cgt atg tat ctt aaa aga gaa gac ttg caa cct gta ttc 144 Leu Gly Val Arg Met Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe 35 40 45 tcg ttt aag ctt cgt gga gct tac aat atg atg gtg aaa ctt cca gca 192 Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala 50 55 60 gat caa ttg gca aaa gga gtt atc tgc tct tca gct gga aac cat gct 240 Asp Gln Leu Ala Lys Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala 65 70 75 80 caa gga gtt gct tta tct gct agt aaa ctc ggc tgc act gct gtg att 288 Gln Gly Val Ala Leu Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile 85 90 95 gtt atg cct gtt acg act cct gag ata aag tgg caa gct gta gag aat 336 Val Met Pro Val Thr Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn 100 105 110 ttg ggt gca acg gtt gtt ctt ttc gga gat tcg tat gat caa gca caa 384 Leu Gly Ala Thr Val Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln 115 120 125 gca cat gct aag ata cga gct gaa gaa gag ggt ctg acg ttt ata cct 432 Ala His Ala Lys Ile Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro 130 135 140 cct ttt gat cac cct gat gtt att gct gga caa ggg act gtt ggg atg 480 Pro Phe Asp His Pro Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met 145 150 155 160 gag atc act cgt cag gct aag ggt cca ttg cat gct ata ttt gtg cca 528 Glu Ile Thr Arg Gln Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro 165 170 175 gtt ggt ggt ggt ggt tta ata gct ggt att gct gct tat gtg aag agg 576 Val Gly Gly Gly Gly Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg 180 185 190 gtt tct ccc gag gtg aag atc att ggt gta gaa cca gct gac gca aat 624 Val Ser Pro Glu Val Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn 195 200 205 gca atg gct ttg tcg ctg cat cac ggt gag agg gtg ata ttg gac cag 672 Ala Met Ala Leu Ser Leu His His Gly Glu Arg Val Ile Leu Asp Gln 210 215 220 gtt ggg gga ttt gca gat ggt gta gca gtt aaa gaa gtt ggt gaa gag 720 Val Gly Gly Phe Ala Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu 225 230 235 240 act ttt cgt ata agc aga aat cta atg gat ggt gtt gtt ctt gtc act 768 Thr Phe Arg Ile Ser Arg Asn Leu Met Asp Gly Val Val Leu Val Thr 245 250 255 cgt gat gct att tgt gca tca ata aag gat atg ttt gag gag aaa cgg 816 Arg Asp Ala Ile Cys Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg 260 265 270 aac ata ttg gaa cca gca ggg gct ctt gca ctc gct gga gct gag gca 864 Asn Ile Leu Glu Pro Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala 275 280 285 tac tgt aaa tat tat ggc cta aag gac gtg aat gtc gta gcc ata acc 912 Tyr Cys Lys Tyr Tyr Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr 290 295 300 agt ggc gct aac atg aac ttt gac aag cta agg att gtg aca gaa ctc 960 Ser Gly Ala Asn Met Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu 305 310 315 320 gcc aat gtc ggt agg caa cag gaa gct gtt ctt gct act ctc atg ccg 1008 Ala Asn Val Gly Arg Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro 325 330 335 gaa aaa cct gga agc ttt aag caa ttt tgt gag ctg gtt gga cca atg 1056 Glu Lys Pro Gly Ser Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met 340 345 350 aac ata agc gag ttc aaa tat aga tgt agc tcg gaa aag gag gct gtt 1104 Asn Ile Ser Glu Phe Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val 355 360 365 gta cta tac agt gtc gga gtt cac aca gct gga gag ctc aaa gca cta 1152 Val Leu Tyr Ser Val Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu 370 375 380 cag aag aga atg gaa tct tct caa ctc aaa act gtc aat ctc act acc 1200 Gln Lys Arg Met Glu Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr 385 390 395 400 agt gac tta gtg aaa gat cac ctg tgt tac ttg atg gga gga aga tct 1248 Ser Asp Leu Val Lys Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser 405 410 415 act gtt gga gac gag gtt cta tgc cga ttc acc ttt ccc gag aga cct 1296 Thr Val Gly Asp Glu Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro 420 425 430 ggt gct cta atg aac ttc ttg gac tct ttc agt cca cgg tgg aac atc 1344 Gly Ala Leu Met Asn Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile 435 440 445 acc ctt ttc cat tac cat gga cag ggt gag acg ggc gcg aat gtg ctg 1392 Thr Leu Phe His Tyr His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu 450 455 460 gtc ggg atc caa gtc ccc gag caa gaa atg gag gaa ttt aaa aac cga 1440 Val Gly Ile Gln Val Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg 465 470 475 480 gct aaa gct ctt gga tac gac tac ttc tta gta agt gat gac gac tat 1488 Ala Lys Ala Leu Gly Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr 485 490 495 ttt aag ctt ctg atg cac tga 1509 Phe Lys Leu Leu Met His 500 8 502 PRT Arabidopsis thaliana 8 Glu Ala Met Glu Tyr Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp 1 5 10 15 Ile Ala Ile Glu Ser Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg 20 25 30 Leu Gly Val Arg Met Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe 35 40 45 Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala 50 55 60 Asp Gln Leu Ala Lys Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala 65 70 75 80 Gln Gly Val Ala Leu Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile 85 90 95 Val Met Pro Val Thr Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn 100 105 110 Leu Gly Ala Thr Val Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln 115 120 125 Ala His Ala Lys Ile Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro 130 135 140 Pro Phe Asp His Pro Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met 145 150 155 160 Glu Ile Thr Arg Gln Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro 165 170 175 Val Gly Gly Gly Gly Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg 180 185 190 Val Ser Pro Glu Val Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn 195 200 205 Ala Met Ala Leu Ser Leu His His Gly Glu Arg Val Ile Leu Asp Gln 210 215 220 Val Gly Gly Phe Ala Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu 225 230 235 240 Thr Phe Arg Ile Ser Arg Asn Leu Met Asp Gly Val Val Leu Val Thr 245 250 255 Arg Asp Ala Ile Cys Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg 260 265 270 Asn Ile Leu Glu Pro Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala 275 280 285 Tyr Cys Lys Tyr Tyr Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr 290 295 300 Ser Gly Ala Asn Met Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu 305 310 315 320 Ala Asn Val Gly Arg Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro 325 330 335 Glu Lys Pro Gly Ser Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met 340 345 350 Asn Ile Ser Glu Phe Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val 355 360 365 Val Leu Tyr Ser Val Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu 370 375 380 Gln Lys Arg Met Glu Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr 385 390 395 400 Ser Asp Leu Val Lys Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser 405 410 415 Thr Val Gly Asp Glu Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro 420 425 430 Gly Ala Leu Met Asn Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile 435 440 445 Thr Leu Phe His Tyr His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu 450 455 460 Val Gly Ile Gln Val Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg 465 470 475 480 Ala Lys Ala Leu Gly Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr 485 490 495 Phe Lys Leu Leu Met His 500 9 1620 DNA Arabidopsis thaliana CDS (1)..(1620) 9 aag ctt cct tta cca cgt ctt aag gtc tct ccg aat tcg ttg caa tac 48 Lys Leu Pro Leu Pro Arg Leu Lys Val Ser Pro Asn Ser Leu Gln Tyr 1 5 10 15 cct gcc ggt tac ctc ggt gct gta cca gaa cgt acg aac gag gct gag 96 Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu Arg Thr Asn Glu Ala Glu 20 25 30 aac gga agc atc gcg gaa gct atg gag tat ttg acg aat ata ctg tcc 144 Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr Leu Thr Asn Ile Leu Ser 35 40 45 act aag gtt tac gac atc gcc att gag tca cca ctc caa ttg gct aag 192 Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser Pro Leu Gln Leu Ala Lys 50 55 60 aag cta tct aag aga tta ggt gtt cgt atg tat ctt aaa aga gaa gac 240 Lys Leu Ser Lys Arg Leu Gly Val Arg Met Tyr Leu Lys Arg Glu Asp 65 70 75 80 ttg caa cct gta ttc tcg ttt aag ctt cgt gga gct tac aat atg atg 288 Leu Gln Pro Val Phe Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Met 85 90 95 gtg aaa ctt cca gca gat caa ttg gca aaa gga gtt atc tgc tct tca 336 Val Lys Leu Pro Ala Asp Gln Leu Ala Lys Gly Val Ile Cys Ser Ser 100 105 110 gct gga aac cat gct caa gga gtt gct tta tct gct agt aaa ctc ggc 384 Ala Gly Asn His Ala Gln Gly Val Ala Leu Ser Ala Ser Lys Leu Gly 115 120 125 tgc act gct gtg att gtt atg cct gtt acg act cct gag ata aag tgg 432 Cys Thr Ala Val Ile Val Met Pro Val Thr Thr Pro Glu Ile Lys Trp 130 135 140 caa gct gta gag aat ttg ggt gca acg gtt gtt ctt ttc gga gat tcg 480 Gln Ala Val Glu Asn Leu Gly Ala Thr Val Val Leu Phe Gly Asp Ser 145 150 155 160 tat gat caa gca caa gca cat gct aag ata cga gct gaa gaa gag ggt 528 Tyr Asp Gln Ala Gln Ala His Ala Lys Ile Arg Ala Glu Glu Glu Gly 165 170 175 ctg acg ttt ata cct cct ttt gat cac cct gat gtt att gct gga caa 576 Leu Thr Phe Ile Pro Pro Phe Asp His Pro Asp Val Ile Ala Gly Gln 180 185 190 ggg act gtt ggg atg gag atc act cgt cag gct aag ggt cca ttg cat 624 Gly Thr Val Gly Met Glu Ile Thr Arg Gln Ala Lys Gly Pro Leu His 195 200 205 gct ata ttt gtg cca gtt ggt ggt ggt ggt tta ata gct ggt att gct 672 Ala Ile Phe Val Pro Val Gly Gly Gly Gly Leu Ile Ala Gly Ile Ala 210 215 220 gct tat gtg aag agg gtt tct ccc gag gtg aag atc att ggt gta gaa 720 Ala Tyr Val Lys Arg Val Ser Pro Glu Val Lys Ile Ile Gly Val Glu 225 230 235 240 cca gct gac gca aat gca atg gct ttg tcg ctg cat cac ggt gag agg 768 Pro Ala Asp Ala Asn Ala Met Ala Leu Ser Leu His His Gly Glu Arg 245 250 255 gtg ata ttg gac cag gtt ggg gga ttt gca gat ggt gta gca gtt aaa 816 Val Ile Leu Asp Gln Val Gly Gly Phe Ala Asp Gly Val Ala Val Lys 260 265 270 gaa gtt ggt gaa gag act ttt cgt ata agc aga aat cta atg gat ggt 864 Glu Val Gly Glu Glu Thr Phe Arg Ile Ser Arg Asn Leu Met Asp Gly 275 280 285 gtt gtt ctt gtc act cgt gat gct att tgt gca tca ata aag gat atg 912 Val Val Leu Val Thr Arg Asp Ala Ile Cys Ala Ser Ile Lys Asp Met 290 295 300 ttt gag gag aaa cgg aac ata ttg gaa cca gca ggg gct ctt gca ctc 960 Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala Gly Ala Leu Ala Leu 305 310 315 320 gct gga gct gag gca tac tgt aaa tat tat ggc cta aag gac gtg aat 1008 Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly Leu Lys Asp Val Asn 325 330 335 gtc gta gcc ata acc agt ggc gct aac atg aac ttt gac aag cta agg 1056 Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn Phe Asp Lys Leu Arg 340 345 350 att gtg aca gaa ctc gcc aat gtc ggt agg caa cag gaa gct gtt ctt 1104 Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln Gln Glu Ala Val Leu 355 360 365 gct act ctc atg ccg gaa aaa cct gga agc ttt aag caa ttt tgt gag 1152 Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe Lys Gln Phe Cys Glu 370 375 380 ctg gtt gga cca atg aac ata agc gag ttc aaa tat aga tgt agc tcg 1200 Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys Tyr Arg Cys Ser Ser 385 390 395 400 gaa aag gag gct gtt gta cta tac agt gtc gga gtt cac aca gct gga 1248 Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly Val His Thr Ala Gly 405 410 415 gag ctc aaa gca cta cag aag aga atg gaa tct tct caa ctc aaa act 1296 Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser Ser Gln Leu Lys Thr 420 425 430 gtc aat ctc act acc agt gac tta gtg aaa gat cac ctg tgt tac ttg 1344 Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp His Leu Cys Tyr Leu 435 440 445 atg gga gga aga tct act gtt gga gac gag gtt cta tgc cga ttc acc 1392 Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val Leu Cys Arg Phe Thr 450 455 460 ttt ccc gag aga cct ggt gct cta atg aac ttc ttg gac tct ttc agt 1440 Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe Leu Asp Ser Phe Ser 465 470 475 480 cca cgg tgg aac atc acc ctt ttc cat tac cat gga cag ggt gag acg 1488 Pro Arg Trp Asn Ile Thr Leu Phe His Tyr His Gly Gln Gly Glu Thr 485 490 495 ggc gcg aat gtg ctg gtc ggg atc caa gtc ccc gag caa gaa atg gag 1536 Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro Glu Gln Glu Met Glu 500 505 510 gaa ttt aaa aac cga gct aaa gct ctt gga tac gac tac ttc tta gta 1584 Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr Asp Tyr Phe Leu Val 515 520 525 agt gat gac gac tat ttt aag ctt ctg atg cac tga 1620 Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 530 535 10 539 PRT Arabidopsis thaliana 10 Lys Leu Pro Leu Pro Arg Leu Lys Val Ser Pro Asn Ser Leu Gln Tyr 1 5 10 15 Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu Arg Thr Asn Glu Ala Glu 20 25 30 Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr Leu Thr Asn Ile Leu Ser 35 40 45 Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser Pro Leu Gln Leu Ala Lys 50 55 60 Lys Leu Ser Lys Arg Leu Gly Val Arg Met Tyr Leu Lys Arg Glu Asp 65 70 75 80 Leu Gln Pro Val Phe Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Met 85 90 95 Val Lys Leu Pro Ala Asp Gln Leu Ala Lys Gly Val Ile Cys Ser Ser 100 105 110 Ala Gly Asn His Ala Gln Gly Val Ala Leu Ser Ala Ser Lys Leu Gly 115 120 125 Cys Thr Ala Val Ile Val Met Pro Val Thr Thr Pro Glu Ile Lys Trp 130 135 140 Gln Ala Val Glu Asn Leu Gly Ala Thr Val Val Leu Phe Gly Asp Ser 145 150 155 160 Tyr Asp Gln Ala Gln Ala His Ala Lys Ile Arg Ala Glu Glu Glu Gly 165 170 175 Leu Thr Phe Ile Pro Pro Phe Asp His Pro Asp Val Ile Ala Gly Gln 180 185 190 Gly Thr Val Gly Met Glu Ile Thr Arg Gln Ala Lys Gly Pro Leu His 195 200 205 Ala Ile Phe Val Pro Val Gly Gly Gly Gly Leu Ile Ala Gly Ile Ala 210 215 220 Ala Tyr Val Lys Arg Val Ser Pro Glu Val Lys Ile Ile Gly Val Glu 225 230 235 240 Pro Ala Asp Ala Asn Ala Met Ala Leu Ser Leu His His Gly Glu Arg 245 250 255 Val Ile Leu Asp Gln Val Gly Gly Phe Ala Asp Gly Val Ala Val Lys 260 265 270 Glu Val Gly Glu Glu Thr Phe Arg Ile Ser Arg Asn Leu Met Asp Gly 275 280 285 Val Val Leu Val Thr Arg Asp Ala Ile Cys Ala Ser Ile Lys Asp Met 290 295 300 Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala Gly Ala Leu Ala Leu 305 310 315 320 Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly Leu Lys Asp Val Asn 325 330 335 Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn Phe Asp Lys Leu Arg 340 345 350 Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln Gln Glu Ala Val Leu 355 360 365 Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe Lys Gln Phe Cys Glu 370 375 380 Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys Tyr Arg Cys Ser Ser 385 390 395 400 Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly Val His Thr Ala Gly 405 410 415 Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser Ser Gln Leu Lys Thr 420 425 430 Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp His Leu Cys Tyr Leu 435 440 445 Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val Leu Cys Arg Phe Thr 450 455 460 Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe Leu Asp Ser Phe Ser 465 470 475 480 Pro Arg Trp Asn Ile Thr Leu Phe His Tyr His Gly Gln Gly Glu Thr 485 490 495 Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro Glu Gln Glu Met Glu 500 505 510 Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr Asp Tyr Phe Leu Val 515 520 525 Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 530 535 11 1599 DNA Arabidopsis thaliana CDS (1)..(1599) 11 aag gtc tct ccg aat tcg ttg caa tac cct gcc ggt tac ctc ggt gct 48 Lys Val Ser Pro Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala 1 5 10 15 gta cca gaa cgt acg aac gag gct gag aac gga agc atc gcg gaa gct 96 Val Pro Glu Arg Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala 20 25 30 atg gag tat ttg acg aat ata ctg tcc act aag gtt tac gac atc gcc 144 Met Glu Tyr Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala 35 40 45 att gag tca cca ctc caa ttg gct aag aag cta tct aag aga tta ggt 192 Ile Glu Ser Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly 50 55 60 gtt cgt atg tat ctt aaa aga gaa gac ttg caa cct gta ttc tcg ttt 240 Val Arg Met Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe 65 70 75 80 aag ctt cgt gga gct tac aat atg atg gtg aaa ctt cca gca gat caa 288 Lys Leu Arg Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln 85 90 95 ttg gca aaa gga gtt atc tgc tct tca gct gga aac cat gct caa gga 336 Leu Ala Lys Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly 100 105 110 gtt gct tta tct gct agt aaa ctc ggc tgc act gct gtg att gtt atg 384 Val Ala Leu Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met 115 120 125 cct gtt acg act cct gag ata aag tgg caa gct gta gag aat ttg ggt 432 Pro Val Thr Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly 130 135 140 gca acg gtt gtt ctt ttc gga gat tcg tat gat caa gca caa gca cat 480 Ala Thr Val Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His 145 150 155 160 gct aag ata cga gct gaa gaa gag ggt ctg acg ttt ata cct cct ttt 528 Ala Lys Ile Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe 165 170 175 gat cac cct gat gtt att gct gga caa ggg act gtt ggg atg gag atc 576 Asp His Pro Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile 180 185 190 act cgt cag gct aag ggt cca ttg cat gct ata ttt gtg cca gtt ggt 624 Thr Arg Gln Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly 195 200 205 ggt ggt ggt tta ata gct ggt att gct gct tat gtg aag agg gtt tct 672 Gly Gly Gly Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser 210 215 220 ccc gag gtg aag atc att ggt gta gaa cca gct gac gca aat gca atg 720 Pro Glu Val Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met 225 230 235 240 gct ttg tcg ctg cat cac ggt gag agg gtg ata ttg gac cag gtt ggg 768 Ala Leu Ser Leu His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly 245 250 255 gga ttt gca gat ggt gta gca gtt aaa gaa gtt ggt gaa gag act ttt 816 Gly Phe Ala Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe 260 265 270 cgt ata agc aga aat cta atg gat ggt gtt gtt ctt gtc act cgt gat 864 Arg Ile Ser Arg Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp 275 280 285 gct att tgt gca tca ata aag gat atg ttt gag gag aaa cgg aac ata 912 Ala Ile Cys Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile 290 295 300 ttg gaa cca gca ggg gct ctt gca ctc gct gga gct gag gca tac tgt 960 Leu Glu Pro Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys 305 310 315 320 aaa tat tat ggc cta aag gac gtg aat gtc gta gcc ata acc agt ggc 1008 Lys Tyr Tyr Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly 325 330 335 gct aac atg aac ttt gac aag cta agg att gtg aca gaa ctc gcc aat 1056 Ala Asn Met Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn 340 345 350 gtc ggt agg caa cag gaa gct gtt ctt gct act ctc atg ccg gaa aaa 1104 Val Gly Arg Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys 355 360 365 cct gga agc ttt aag caa ttt tgt gag ctg gtt gga cca atg aac ata 1152 Pro Gly Ser Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile 370 375 380 agc gag ttc aaa tat aga tgt agc tcg gaa aag gag gct gtt gta cta 1200 Ser Glu Phe Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu 385 390 395 400 tac agt gtc gga gtt cac aca gct gga gag ctc aaa gca cta cag aag 1248 Tyr Ser Val Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys 405 410 415 aga atg gaa tct tct caa ctc aaa act gtc aat ctc act acc agt gac 1296 Arg Met Glu Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp 420 425 430 tta gtg aaa gat cac ctg tgt tac ttg atg gga gga aga tct act gtt 1344 Leu Val Lys Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val 435 440 445 gga gac gag gtt cta tgc cga ttc acc ttt ccc gag aga cct ggt gct 1392 Gly Asp Glu Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala 450 455 460 cta atg aac ttc ttg gac tct ttc agt cca cgg tgg aac atc acc ctt 1440 Leu Met Asn Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu 465 470 475 480 ttc cat tac cat gga cag ggt gag acg ggc gcg aat gtg ctg gtc ggg 1488 Phe His Tyr His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly 485 490 495 atc caa gtc ccc gag caa gaa atg gag gaa ttt aaa aac cga gct aaa 1536 Ile Gln Val Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys 500 505 510 gct ctt gga tac gac tac ttc tta gta agt gat gac gac tat ttt aag 1584 Ala Leu Gly Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys 515 520 525 ctt ctg atg cac tga 1599 Leu Leu Met His 530 12 532 PRT Arabidopsis thaliana 12 Lys Val Ser Pro Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala 1 5 10 15 Val Pro Glu Arg Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala 20 25 30 Met Glu Tyr Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala 35 40 45 Ile Glu Ser Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly 50 55 60 Val Arg Met Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe 65 70 75 80 Lys Leu Arg Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln 85 90 95 Leu Ala Lys Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly 100 105 110 Val Ala Leu Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met 115 120 125 Pro Val Thr Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly 130 135 140 Ala Thr Val Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His 145 150 155 160 Ala Lys Ile Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe 165 170 175 Asp His Pro Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile 180 185 190 Thr Arg Gln Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly 195 200 205 Gly Gly Gly Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser 210 215 220 Pro Glu Val Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met 225 230 235 240 Ala Leu Ser Leu His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly 245 250 255 Gly Phe Ala Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe 260 265 270 Arg Ile Ser Arg Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp 275 280 285 Ala Ile Cys Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile 290 295 300 Leu Glu Pro Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys 305 310 315 320 Lys Tyr Tyr Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly 325 330 335 Ala Asn Met Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn 340 345 350 Val Gly Arg Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys 355 360 365 Pro Gly Ser Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile 370 375 380 Ser Glu Phe Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu 385 390 395 400 Tyr Ser Val Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys 405 410 415 Arg Met Glu Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp 420 425 430 Leu Val Lys Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val 435 440 445 Gly Asp Glu Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala 450 455 460 Leu Met Asn Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu 465 470 475 480 Phe His Tyr His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly 485 490 495 Ile Gln Val Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys 500 505 510 Ala Leu Gly Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys 515 520 525 Leu Leu Met His 530 13 720 DNA Arabidopsis thaliana CDS (1)..(720) 13 tca ata aag gat atg ttt gag gag aaa cgg aac ata ttg gaa cca gca 48 Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala 1 5 10 15 ggg gct ctt gca ctc gct gga gct gag gca tac tgt aaa tat tat ggc 96 Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly 20 25 30 cta aag gac gtg aat gtc gta gcc ata acc agt ggc gct aac atg aac 144 Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn 35 40 45 ttt gac aag cta agg att gtg aca gaa ctc gcc aat gtc ggt agg caa 192 Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln 50 55 60 cag gaa gct gtt ctt gct act ctc atg ccg gaa aaa cct gga agc ttt 240 Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe 65 70 75 80 aag caa ttt tgt gag ctg gtt gga cca atg aac ata agc gag ttc aaa 288 Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys 85 90 95 tat aga tgt agc tcg gaa aag gag gct gtt gta cta tac agt gtc gga 336 Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly 100 105 110 gtt cac aca gct gga gag ctc aaa gca cta cag aag aga atg gaa tct 384 Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser 115 120 125 tct caa ctc aaa act gtc aat ctc act acc agt gac tta gtg aaa gat 432 Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp 130 135 140 cac ctg tgt tac ttg atg gga gga aga tct act gtt gga gac gag gtt 480 His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 145 150 155 160 cta tgc cga ttc acc ttt ccc gag aga cct ggt gct cta atg aac ttc 528 Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe 165 170 175 ttg gac tct ttc agt cca cgg tgg aac atc acc ctt ttc cat tac cat 576 Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr His 180 185 190 gga cag ggt gag acg ggc gcg aat gtg ctg gtc ggg atc caa gtc ccc 624 Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro 195 200 205 gag caa gaa atg gag gaa ttt aaa aac cga gct aaa gct ctt gga tac 672 Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr 210 215 220 gac tac ttc tta gta agt gat gac gac tat ttt aag ctt ctg atg cac 720 Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 225 230 235 240 14 240 PRT Arabidopsis thaliana 14 Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala 1 5 10 15 Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly 20 25 30 Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn 35 40 45 Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln 50 55 60 Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe 65 70 75 80 Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys 85 90 95 Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly 100 105 110 Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser 115 120 125 Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp 130 135 140 His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 145 150 155 160 Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe 165 170 175 Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr His 180 185 190 Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro 195 200 205 Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr 210 215 220 Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 225 230 235 240 15 81 DNA Arabidopsis thaliana CDS (1)..(81) 15 gtc aat ctc act acc agt gac tta gtg aaa gat cac ctg tgt tac ttg 48 Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp His Leu Cys Tyr Leu 1 5 10 15 atg gga gga aga tct act gtt gga gac gag gtt 81 Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 20 25 16 27 PRT Arabidopsis thaliana 16 Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp His Leu Cys Tyr Leu 1 5 10 15 Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 20 25 17 75 DNA Arabidopsis thaliana CDS (1)..(75) 17 tgg aac atc acc ctt ttc cat tac cat gga cag ggt gag acg ggc gcg 48 Trp Asn Ile Thr Leu Phe His Tyr His Gly Gln Gly Glu Thr Gly Ala 1 5 10 15 aat gtg ctg gtc ggg atc caa gtc ccc 75 Asn Val Leu Val Gly Ile Gln Val Pro 20 25 18 25 PRT Arabidopsis thaliana 18 Trp Asn Ile Thr Leu Phe His Tyr His Gly Gln Gly Glu Thr Gly Ala 1 5 10 15 Asn Val Leu Val Gly Ile Gln Val Pro 20 25 19 1638 DNA Arabidopsis thaliana CDS (1)..(1638) 19 atg act cca ccg cct cca aag ctt cct tta cca cgt ctt aag gtc tct 48 Met Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser 1 5 10 15 ccg aat tcg ttg caa tac cct gcc ggt tac ctc ggt gct gta cca gaa 96 Pro Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu 20 25 30 cgt acg aac gag gct gag aac gga agc atc gcg gaa gct atg gag tat 144 Arg Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr 35 40 45 ttg acg aat ata ctg tcc act aag gtt tac gac atc gcc att gag tca 192 Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser 50 55 60 cca ctc caa ttg gct aag aag cta tct aag aga tta ggt gtt cgt atg 240 Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met 65 70 75 80 tat ctt aaa aga gaa gac ttg caa cct gta ttc tcg ttt aag ctt cgt 288 Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg 85 90 95 gga gct tac aat atg atg gtg aaa ctt cca gca gat caa ttg gca aaa 336 Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys 100 105 110 gga gtt atc tgc tct tca gct gga aac cat gct caa gga gtt gct tta 384 Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu 115 120 125 tct gct agt aaa ctc ggc tgc act gct gtg att gtt atg cct gtt acg 432 Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr 130 135 140 act cct gag ata aag tgg caa gct gta gag aat ttg ggt gca acg gtt 480 Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val 145 150 155 160 gtt ctt ttc gga gat tcg tat gat caa gca caa gca cat gct aag ata 528 Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile 165 170 175 cga gct gaa gaa gag ggt ctg acg ttt ata cct cct ttt gat cac cct 576 Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro 180 185 190 gat gtt att gct gga caa ggg act gtt ggg atg gag atc act cgt cag 624 Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln 195 200 205 gct aag ggt cca ttg cat gct ata ttt gtg cca gtt ggt ggt ggt ggt 672 Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly 210 215 220 tta ata gct ggt att gct gct tat gtg aag agg gtt tct ccc gag gtg 720 Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val 225 230 235 240 aag atc att ggt gta gaa cca gct gac gca aat gca atg gct ttg tcg 768 Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser 245 250 255 ctg cat cac ggt gag agg gtg ata ttg gac cag gtt ggg gga ttt gca 816 Leu His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala 260 265 270 gat ggt gta gca gtt aaa gaa gtt ggt gaa gag act ttt cgt ata agc 864 Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser 275 280 285 aga aat cta atg gat ggt gtt gtt ctt gtc act cgt gat gct att tgt 912 Arg Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys 290 295 300 gca tca ata aag gat atg ttt gag gag aaa cgg aac ata ttg gaa cca 960 Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro 305 310 315 320 gca ggg gct ctt gca ctc gct gga gct gag gca tac tgt aaa tat tat 1008 Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr 325 330 335 ggc cta aag gac gtg aat gtc gta gcc ata acc agt ggc gct aac atg 1056 Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met 340 345 350 aac ttt gac aag cta agg att gtg aca gaa ctc gcc aat gtc ggt agg 1104 Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg 355 360 365 caa cag gaa gct gtt ctt gct act ctc atg ccg gaa aaa cct gga agc 1152 Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser 370 375 380 ttt aag caa ttt tgt gag ctg gtt gga cca atg aac ata agc gag ttc 1200 Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe 385 390 395 400 aaa tat aga tgt agc tcg gaa aag gag gct gtt gta cta tac agt gtc 1248 Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val 405 410 415 gga gtt cac aca gct gga gag ctc aaa gca cta cag aag aga atg gaa 1296 Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu 420 425 430 tct tct caa ctc aaa act gtc aat ctc act acc agt gac tta gtg aaa 1344 Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys 435 440 445 gat cac ctg tgt tac ttg atg gga gga aga tct act gtt gga gac gag 1392 Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu 450 455 460 gtt cta tgc cga ttc acc ttt ccc gag aga cct ggt gct cta atg aac 1440 Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn 465 470 475 480 ttc ttg gac tct ttc agt cca cgg tgg aac atc acc ctt ttc cat tac 1488 Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr 485 490 495 cat gga cag ggt gag acg ggc gcg aat gtg ctg gtc ggg atc caa gtc 1536 His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val 500 505 510 ccc gag caa gaa atg gag gaa ttt aaa aac cga gct aaa gct ctt gga 1584 Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly 515 520 525 tac gac tac ttc tta gta agt gat gac gac tat ttt aag ctt ctg atg 1632 Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met 530 535 540 cac tga 1638 His 545 20 545 PRT Arabidopsis thaliana 20 Met Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser 1 5 10 15 Pro Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu 20 25 30 Arg Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr 35 40 45 Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser 50 55 60 Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met 65 70 75 80 Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg 85 90 95 Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys 100 105 110 Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu 115 120 125 Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr 130 135 140 Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val 145 150 155 160 Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile 165 170 175 Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro 180 185 190 Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln 195 200 205 Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly 210 215 220 Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val 225 230 235 240 Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser 245 250 255 Leu His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala 260 265 270 Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser 275 280 285 Arg Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys 290 295 300 Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro 305 310 315 320 Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr 325 330 335 Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met 340 345 350 Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg 355 360 365 Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser 370 375 380 Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe 385 390 395 400 Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val 405 410 415 Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu 420 425 430 Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys 435 440 445 Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu 450 455 460 Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn 465 470 475 480 Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr 485 490 495 His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val 500 505 510 Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly 515 520 525 Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met 530 535 540 His 545 21 19 PRT Arabidopsis thaliana 21 Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp His Leu Arg Tyr Leu 1 5 10 15 Met Gly Gly 22 19 PRT Artificial Sequence Synthetic 22 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Gly Gly 23 19 PRT Artificial Sequence Synthetic 23 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa His Xaa Cys Xaa Xaa 1 5 10 15 Xaa Gly Gly 24 19 PRT Arabidopsis thaliana 24 Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp His Leu Cys Tyr Leu 1 5 10 15 Met Gly Gly 25 19 PRT Arabidopsis thaliana 25 Trp Asn Ile Thr Leu Phe His Tyr His Gly Gln Gly Glu Thr Gly Ala 1 5 10 15 Asn Val Leu 26 590 PRT Cicer arietinum 26 Met Leu Ser Thr Ser Thr Thr Asn Ser Ser Ile Leu Pro Phe Arg Ser 1 5 10 15 Arg Ala Ser Ser Ser Thr Phe Ile Ala Arg Pro Pro Ala Asn Phe Asn 20 25 30 Ser Ile Phe Thr Thr Ser Val Arg Val Phe Pro Ile Ser Met Ser Arg 35 40 45 Tyr Cys Val Phe Pro His Thr Trp Glu Arg Asp His Asn Val Pro Gly 50 55 60 Val Pro Gly Val Leu Arg Lys Val Val Pro Ala Ala Pro Ile Lys Asn 65 70 75 80 Lys Pro Thr Cys Ala Asp Ser Asp Glu Leu Pro Glu Tyr Leu Arg Asp 85 90 95 Val Leu Arg Ser Pro Val Tyr Asp Val Val Val Glu Ser Pro Val Glu 100 105 110 Leu Thr Glu Arg Leu Ser Asp Arg Leu Gly Val Asn Phe Tyr Val Lys 115 120 125 Arg Glu Asp Arg Gln Arg Val Phe Ser Phe Lys Leu Arg Gly Pro Tyr 130 135 140 Asn Met Met Ser Ser Leu Ser His Glu Glu Ile Asp Lys Gly Val Ile 145 150 155 160 Thr Ala Ser Ala Gly Asn His Ala Gln Gly Val Pro Phe Pro Phe Pro 165 170 175 Gly Arg Arg Leu Lys Cys Val Ala Lys Ile Val Met Pro Thr Thr Thr 180 185 190 Pro Asn Ile Lys Leu Asp Gly Val Arg Ala Leu Gly Ala Asp Val Val 195 200 205 Leu Trp Gly His Thr Phe Asp Glu Ala Lys Thr His Ala Val Glu Leu 210 215 220 Cys Glu Lys Asp Gly Leu Arg Thr Ile Pro Pro Phe Glu Asp Pro Ala 225 230 235 240 Val Ile Lys Gly Gln Gly Thr Ile Gly Ser Glu Ile Asn Arg Gln Ile 245 250 255 Lys Arg Ile Asp Ala Val Phe Val Pro Val Gly Gly Gly Gly Leu Ile 260 265 270 Ala Gly Val Ala Ala Phe Phe Lys Gln Ile Ala Pro Gln Thr Lys Ile 275 280 285 Ile Val Val Glu Pro Tyr Asp Ala Ala Ser Met Ala Leu Ser Val His 290 295 300 Ala Glu His Arg Ala Lys Leu Ser Asn Val Asp Thr Phe Ala Asp Gly 305 310 315 320 Ala Thr Val Ala Val Ile Gly Glu Tyr Thr Phe Ala Arg Cys Gln Asp 325 330 335 Val Val Asp Ala Met Val Leu Val Ala Asn Asp Gly Ile Gly Ala Ala 340 345 350 Ile Lys Asp Val Phe Asp Glu Gly Arg Asn Ile Val Glu Thr Ser Gly 355 360 365 Ala Ala Gly Ile Ala Gly Met Tyr Cys Glu Met Tyr Arg Ile Lys Asn 370 375 380 Asp Asn Met Val Gly Ile Val Ser Gly Ala Asn Met Asn Phe Arg Lys 385 390 395 400 Leu His Lys Val Ser Glu Leu Ala Val Leu Gly Ser Gly His Glu Ala 405 410 415 Leu Leu Gly Thr Tyr Met Pro Gly Gln Lys Gly Cys Phe Lys Thr Met 420 425 430 Ala Gly Leu Val His Gly Ser Leu Ser Phe Thr Glu Ile Thr Tyr Arg 435 440 445 Phe Thr Ser His Arg Arg Ser Ile Leu Val Leu Met Leu Lys Leu Glu 450 455 460 Pro Trp Arg Tyr Ile Glu Lys Met Ile Glu Met Met Lys Tyr Ser Gly 465 470 475 480 Val Thr Val Leu Asn Ile Ser His Asn Glu Leu Ala Val Ile His Gly 485 490 495 Lys His Leu Val Gly Gly Ser Ala Lys Val Ser Asp Glu Val Phe Val 500 505 510 Glu Phe Ile Ile Pro Glu Lys Ala Asp Leu Lys Lys Phe Leu Glu Val 515 520 525 Leu Ser Pro His Trp Asn Leu Thr Leu Tyr Arg Tyr Arg Asn Gln Gly 530 535 540 Asp Leu Lys Ala Thr Ile Leu Met Val Ile Ala Ser Phe Leu Cys Glu 545 550 555 560 Ile Val Ile Arg Lys Asn Gln Ile Asp Asp Leu Gly Tyr Pro Tyr Glu 565 570 575 Ile Asp Gln Tyr Asn Asp Ala Phe Asn Leu Ala Val Thr Glu 580 585 590 27 595 PRT Lycopersicon esculentum 27 Met Glu Phe Leu Cys Leu Ala Pro Thr Arg Ser Phe Ser Thr Asn Pro 1 5 10 15 Lys Leu Thr Lys Ser Ile Pro Ser Asp His Thr Ser Thr Thr Ser Arg 20 25 30 Ile Phe Thr Tyr Gln Asn Met Arg Gly Ser Thr Met Arg Pro Leu Ala 35 40 45 Leu Pro Leu Lys Met Ser Pro Ile Val Ser Val Pro Asp Ile Thr Ala 50 55 60 Pro Val Glu Asn Val Pro Ala Ile Leu Pro Lys Val Val Pro Gly Glu 65 70 75 80 Leu Ile Val Asn Lys Pro Thr Gly Gly Asp Ser Asp Glu Leu Phe Gln 85 90 95 Tyr Leu Val Asp Ile Leu Ala Ser Pro Val Tyr Asp Val Ala Ile Glu 100 105 110 Ser Pro Leu Glu Leu Ala Glu Lys Leu Ser Asp Arg Leu Gly Val Asn 115 120 125 Phe Tyr Ile Lys Arg Glu Asp Lys Gln Arg Val Phe Ser Phe Lys Leu 130 135 140 Arg Gly Ala Tyr Asn Met Met Ser Asn Leu Ser Arg Glu Glu Leu Asp 145 150 155 160 Lys Gly Val Ile Thr Ala Ser Ala Gly Asn His Ala Gln Gly Val Ala 165 170 175 Leu Ala Gly Gln Arg Leu Asn Cys Val Ala Lys Ile Val Met Pro Thr 180 185 190 Thr Thr Pro Gln Ile Lys Ile Asp Ala Val Arg Ala Leu Gly Gly Asp 195 200 205 Val Val Leu Tyr Gly Lys Thr Phe Asp Glu Ala Gln Thr His Ala Leu 210 215 220 Glu Leu Ser Glu Lys Asp Gly Leu Lys Tyr Ile Pro Pro Phe Asp Asp 225 230 235 240 Pro Gly Val Ile Lys Gly Gln Gly Thr Ile Gly Thr Glu Ile Asn Arg 245 250 255 Gln Leu Lys Asp Ile His Ala Val Phe Ile Pro Val Gly Gly Gly Gly 260 265 270 Leu Ile Ala Gly Val Ala Thr Phe Phe Lys Gln Ile Ala Pro Asn Thr 275 280 285 Lys Ile Ile Gly Val Glu Pro Tyr Gly Ala Ala Ser Met Thr Leu Ser 290 295 300 Leu His Glu Gly His Arg Val Lys Leu Ser Asn Val Asp Thr Phe Ala 305 310 315 320 Asp Gly Val Ala Val Ala Leu Val Gly Glu Tyr Thr Phe Ala Lys Cys 325 330 335 Gln Glu Leu Ile Asp Gly Met Val Leu Val Ala Asn Asp Gly Ile Ser 340 345 350 Ala Ala Ile Lys Asp Val Tyr Asp Glu Gly Arg Asn Ile Leu Glu Thr 355 360 365 Ser Gly Ala Val Ala Ile Ala Gly Ala Ala Ala Tyr Cys Glu Phe Tyr 370 375 380 Lys Ile Lys Asn Glu Asn Ile Val Ala Ile Ala Ser Gly Ala Asn Met 385 390 395 400 Asp Phe Ser Lys Leu His Lys Val Thr Glu Leu Ala Gly Leu Gly Ser 405 410 415 Gly Lys Glu Ala Leu Leu Ala Thr Phe Met Val Glu Gln Gln Gly Ser 420 425 430 Phe Lys Thr Phe Val Gly Leu Val Gly Ser Leu Asn Phe Thr Glu Leu 435 440 445 Thr Tyr Arg Phe Thr Ser Glu Arg Lys Asn Ala Leu Ile Leu Tyr Arg 450 455 460 Val Asn Val Asp Lys Glu Ser Asp Leu Glu Lys Met Ile Glu Asp Met 465 470 475 480 Lys Ser Ser Asn Met Thr Thr Leu Asn Leu Ser His Asn Glu Leu Val 485 490 495 Val Asp His Leu Lys His Leu Val Gly Gly Ser Ala Asn Ile Ser Asp 500 505 510 Glu Ile Phe Gly Glu Phe Ile Val Pro Glu Lys Ala Glu Thr Leu Lys 515 520 525 Thr Phe Leu Asp Ala Phe Ser Pro Arg Trp Asn Ile Thr Leu Cys Arg 530 535 540 Tyr Arg Asn Gln Gly Asp Ile Asn Ala Ser Leu Leu Met Gly Phe Gln 545 550 555 560 Val Pro Gln Ala Glu Met Asp Glu Phe Lys Asn Gln Ala Asp Lys Leu 565 570 575 Gly Tyr Pro Tyr Glu Leu Asp Asn Tyr Asn Glu Ala Phe Asn Leu Val 580 585 590 Val Ser Glu 595 28 359 PRT Solanum tuberosum 28 Pro Phe Asp Ala Pro Gly Val Ile Lys Gly Gln Gly Thr Ile Gly Thr 1 5 10 15 Glu Ile Asn Arg Gln Leu Lys Asp Ile His Ala Val Phe Val Pro Val 20 25 30 Gly Gly Gly Gly Leu Ile Ser Gly Val Ala Ala Tyr Phe Thr Gln Val 35 40 45 Ala Pro His Thr Lys Ile Ile Gly Val Glu Pro Tyr Gly Ala Ala Ser 50 55 60 Met Thr Leu Ser Leu Tyr Glu Gly His Arg Val Lys Leu Glu Asn Val 65 70 75 80 Asp Thr Phe Ala Asp Gly Val Ala Val Ala Leu Val Gly Glu Tyr Thr 85 90 95 Phe Ala Lys Cys Gln Glu Leu Ile Asp Gly Met Val Leu Val Arg Asn 100 105 110 Asp Gly Ile Ser Ala Ala Ile Lys Asp Val Tyr Asp Glu Gly Arg Asn 115 120 125 Ile Leu Glu Thr Ser Gly Ala Val Ala Ile Ala Gly Ala Ala Ala Tyr 130 135 140 Cys Glu Phe Tyr Asn Ile Lys Asn Glu Asn Ile Val Ala Ile Ala Ser 145 150 155 160 Gly Ala Asn Met Asp Phe Ser Lys Leu His Lys Val Thr Glu Leu Ala 165 170 175 Glu Leu Gly Ser Asp Asn Glu Ala Leu Leu Ala Thr Phe Met Ile Glu 180 185 190 Gln Pro Gly Ser Phe Lys Thr Phe Ala Lys Leu Val Gly Ser Met Asn 195 200 205 Ile Thr Glu Val Thr Tyr Arg Phe Thr Ser Glu Arg Lys Glu Ala Leu 210 215 220 Val Leu Tyr Arg Val Asp Val Asp Glu Lys Ser Asp Leu Glu Glu Met 225 230 235 240 Ile Lys Lys Leu Asn Ser Ser Asn Met Lys Thr Phe Asn Phe Ser His 245 250 255 Asn Glu Leu Val Ala Glu His Ile Lys His Leu Val Gly Gly Ser Ala 260 265 270 Ser Ile Ser Asp Glu Ile Phe Gly Glu Phe Ile Phe Pro Glu Lys Ala 275 280 285 Gly Thr Leu Ser Thr Phe Leu Glu Ala Phe Ser Pro Arg Trp Asn Ile 290 295 300 Thr Leu Cys Arg Tyr Arg Asp Gln Gly Asp Ile Asn Gly Asn Val Leu 305 310 315 320 Val Gly Phe Gln Val Pro Gln Ser Glu Met Asp Glu Phe Lys Ser Gln 325 330 335 Ala Asp Gly Leu Gly Tyr Pro Tyr Glu Leu Asp Asn Ser Asn Glu Ala 340 345 350 Phe Asn Ile Val Val Ala Glu 355 29 576 PRT Saccharomyces cerevisiae 29 Met Ser Ala Thr Leu Leu Lys Gln Pro Leu Cys Thr Val Val Arg Gln 1 5 10 15 Gly Lys Gln Ser Lys Val Ser Gly Leu Asn Leu Leu Arg Leu Lys Ala 20 25 30 His Leu His Arg Gln His Leu Ser Pro Ser Leu Ile Lys Leu His Ser 35 40 45 Glu Leu Lys Leu Asp Glu Leu Gln Thr Asp Asn Thr Pro Asp Tyr Val 50 55 60 Arg Leu Val Leu Arg Ser Ser Val Tyr Asp Val Ile Asn Glu Ser Pro 65 70 75 80 Ile Ser Gln Gly Val Gly Leu Ser Ser Arg Leu Asn Thr Asn Val Ile 85 90 95 Leu Lys Arg Glu Asp Leu Leu Pro Val Phe Ser Phe Lys Leu Arg Gly 100 105 110 Ala Tyr Asn Met Ile Ala Lys Leu Asp Asp Ser Gln Arg Asn Gln Gly 115 120 125 Val Ile Ala Cys Ser Ala Gly Asn His Ala Gln Gly Val Ala Phe Ala 130 135 140 Ala Lys His Leu Lys Ile Pro Ala Thr Ile Val Met Pro Val Cys Thr 145 150 155 160 Pro Ser Ile Lys Tyr Gln Asn Val Ser Arg Leu Gly Ser Gln Val Val 165 170 175 Leu Tyr Gly Asn Asp Phe Asp Glu Ala Lys Ala Glu Cys Ala Lys Leu 180 185 190 Ala Glu Glu Arg Gly Leu Thr Asn Ile Pro Pro Phe Asp His Pro Tyr 195 200 205 Val Ile Ala Gly Gln Gly Thr Val Ala Met Glu Ile Leu Arg Gln Val 210 215 220 Arg Thr Ala Asn Lys Ile Gly Ala Val Phe Val Pro Val Gly Gly Gly 225 230 235 240 Gly Leu Ile Ala Gly Ile Gly Ala Tyr Leu Lys Arg Val Ala Pro His 245 250 255 Ile Lys Thr Ile Gly Val Glu Thr Tyr Asp Ala Ala Thr Leu His Asn 260 265 270 Ser Leu Gln Arg Asn Gln Arg Thr Pro Leu Pro Val Val Gly Thr Phe 275 280 285 Ala Asp Gly Thr Ser Val Arg Met Ile Gly Glu Glu Thr Phe Arg Val 290 295 300 Ala Gln Gln Val Val Asp Glu Val Val Leu Val Asn Thr Asp Glu Ile 305 310 315 320 Cys Ala Ala Val Lys Asp Ile Phe Glu Asp Thr Arg Ser Ile Val Glu 325 330 335 Pro Ser Gly Ala Leu Ser Val Ala Gly Met Lys Lys Tyr Ile Ser Thr 340 345 350 Val His Pro Glu Ile Asp His Thr Lys Asn Thr Tyr Val Pro Ile Leu 355 360 365 Ser Gly Ala Asn Met Asn Phe Asp Arg Leu Arg Phe Val Ser Glu Arg 370 375 380 Ala Val Leu Gly Glu Gly Lys Glu Val Phe Met Leu Val Thr Leu Pro 385 390 395 400 Asp Val Pro Gly Ala Phe Lys Lys Met Gln Lys Ile Ile His Pro Arg 405 410 415 Ser Val Thr Glu Phe Ser Tyr Arg Tyr Asn Glu His Arg His Glu Ser 420 425 430 Ser Ser Glu Val Pro Lys Ala Tyr Ile Tyr Thr Ser Phe Ser Val Val 435 440 445 Asp Arg Glu Lys Glu Ile Lys Gln Val Met Gln Gln Leu Asn Ala Leu 450 455 460 Gly Phe Glu Ala Val Asp Ile Ser Asp Asn Glu Leu Ala Lys Ser His 465 470 475 480 Gly Arg Tyr Leu Val Gly Gly Ala Ser Lys Val Pro Asn Glu Arg Ile 485 490 495 Ile Ser Phe Glu Phe Pro Glu Arg Pro Gly Ala Leu Thr Arg Phe Leu 500 505 510 Gly Gly Leu Ser Asp Ser Trp Asn Leu Thr Leu Phe His Tyr Arg Asn 515 520 525 His Gly Ala Asp Ile Gly Lys Val Leu Ala Gly Ile Ser Val Pro Pro 530 535 540 Arg Glu Asn Leu Thr Phe Gln Lys Phe Leu Glu Asp Leu Gly Tyr Thr 545 550 555 560 Tyr His Asp Glu Thr Asp Asn Thr Val Tyr Gln Lys Phe Leu Lys Tyr 565 570 575 30 360 PRT Saccharomyces cerevisiae 30 Met Ser Ile Val Tyr Asn Lys Thr Pro Leu Leu Arg Gln Phe Phe Pro 1 5 10 15 Gly Lys Ala Ser Ala Gln Phe Phe Leu Lys Tyr Glu Cys Leu Gln Pro 20 25 30 Ser Gly Ser Phe Lys Ser Arg Gly Ile Gly Asn Leu Ile Met Lys Ser 35 40 45 Ala Ile Arg Ile Gln Lys Asp Gly Lys Arg Ser Pro Gln Val Phe Ala 50 55 60 Ser Ser Gly Gly Asn Ala Gly Phe Ala Ala Ala Thr Ala Cys Gln Arg 65 70 75 80 Leu Ser Leu Pro Cys Thr Val Val Val Pro Thr Ala Thr Lys Lys Arg 85 90 95 Met Val Asp Lys Ile Arg Asn Thr Gly Ala Gln Val Ile Val Ser Gly 100 105 110 Ala Tyr Trp Lys Glu Ala Asp Thr Phe Leu Lys Thr Asn Val Met Asn 115 120 125 Lys Ile Asp Ser Gln Val Ile Glu Pro Ile Tyr Val His Pro Phe Asp 130 135 140 Asn Pro Asp Ile Trp Glu Gly His Ser Ser Met Ile Asp Glu Ile Val 145 150 155 160 Gln Asp Leu Lys Ser Gln His Ile Ser Val Asn Lys Val Lys Gly Ile 165 170 175 Val Cys Ser Val Gly Gly Gly Gly Leu Tyr Asn Gly Ile Ile Gln Gly 180 185 190 Leu Glu Arg Tyr Gly Leu Ala Asp Arg Ile Pro Ile Val Gly Val Glu 195 200 205 Thr Asn Gly Cys His Val Phe Asn Thr Ser Leu Lys Ile Gly Gln Pro 210 215 220 Val Gln Phe Lys Lys Ile Thr Ser Ile Ala Thr Ser Leu Gly Thr Ala 225 230 235 240 Val Ile Ser Asn Gln Thr Phe Glu Tyr Ala Arg Lys Tyr Asn Thr Arg 245 250 255 Ser Val Val Ile Glu Asp Lys Asp Val Ile Glu Pro Cys Leu Lys Tyr 260 265 270 Thr His Gln Phe Asn Met Val Ile Glu Pro Ala Cys Gly Ala Ala Leu 275 280 285 His Leu Gly Tyr Asn Thr Lys Ile Leu Glu Asn Ala Leu Gly Ser Lys 290 295 300 Leu Ala Ala Asp Asp Ile Val Ile Ile Ile Ala Cys Ala Ser Ser Ser 305 310 315 320 Asn Thr Ile Lys Asp Leu Glu Glu Ala Leu Asp Ser Met Arg Lys Lys 325 330 335 Asp Thr Pro Val Ile Glu Val Ala Asp Asn Phe Ile Phe Pro Glu Lys 340 345 350 Asn Ile Val Asn Leu Lys Ser Ala 355 360 31 514 PRT Salmonella typhimurium 31 Met Ala Glu Ser Gln Pro Leu Ser Val Ala Pro Glu Gly Ala Glu Tyr 1 5 10 15 Leu Arg Ala Val Leu Arg Ala Pro Val Tyr Glu Ala Ala Gln Val Thr 20 25 30 Pro Leu Gln Lys Met Glu Lys Leu Ser Ser Arg Leu Asp Asn Val Ile 35 40 45 Leu Val Lys Arg Glu Asp Arg Gln Pro Val His Ser Phe Lys Leu Arg 50 55 60 Gly Ala Tyr Ala Met Met Ala Gly Leu Thr Glu Glu Gln Lys Ala His 65 70 75 80 Gly Val Ile Thr Ala Ser Ala Gly Asn His Ala Gln Gly Val Ala Phe 85 90 95 Ser Ser Ala Arg Leu Gly Val Lys Ser Leu Ile Val Met Pro Lys Ala 100 105 110 Thr Ala Asp Ile Lys Val Asp Ala Val Arg Gly Phe Gly Gly Glu Val 115 120 125 Leu Leu His Gly Ala Asn Phe Asp Glu Ala Lys Ala Lys Ala Ile Glu 130 135 140 Leu Ala Gln Gln Gln Gly Phe Thr Trp Val Pro Pro Phe Asp His Pro 145 150 155 160 Met Val Ile Ala Gly Gln Gly Thr Leu Ala Leu Glu Leu Leu Gln Gln 165 170 175 Asp Ser His Leu Asp Arg Val Phe Val Pro Val Gly Gly Gly Gly Leu 180 185 190 Ala Ala Gly Val Ala Val Leu Ile Lys Gln Leu Met Pro Gln Ile Lys 195 200 205 Val Ile Ala Val Glu Ala Glu Asp Ser Ala Cys Leu Lys Ala Ala Leu 210 215 220 Glu Ala Gly His Pro Val Asp Leu Pro Arg Val Gly Leu Phe Ala Glu 225 230 235 240 Gly Val Ala Val Lys Arg Ile Gly Asp Glu Thr Phe Arg Leu Cys Gln 245 250 255 Glu Tyr Leu Asp Asp Ile Ile Thr Val Asp Ser Asp Ala Ile Cys Ala 260 265 270 Ala Met Lys Asp Leu Phe Glu Asp Val Arg Ala Val Ala Glu Pro Ser 275 280 285 Gly Ala Leu Ala Leu Ala Gly Met Lys Lys Tyr Ile Ala Gln His Asn 290 295 300 Ile Arg Gly Glu Arg Leu Ala His Val Leu Ser Gly Ala Asn Val Asn 305 310 315 320 Phe His Gly Leu Arg Tyr Val Ser Glu Arg Cys Glu Leu Gly Glu Gln 325 330 335 Arg Glu Ala Leu Leu Ala Val Thr Ile Pro Glu Glu Lys Gly Ser Phe 340 345 350 Leu Lys Phe Cys Gln Leu Leu Gly Gly Arg Met Val Thr Glu Phe Asn 355 360 365 Tyr Arg Phe Ala Asp Ala Lys Asn Ala Cys Ile Phe Val Gly Val Arg 370 375 380 Val Ser Gln Gly Leu Glu Glu Arg Lys Glu Ile Ile Thr Gln Leu Cys 385 390 395 400 Asp Gly Gly Tyr Ser Val Val Asp Leu Ser Asp Asp Glu Met Ala Lys 405 410 415 Leu His Val Arg Tyr Met Val Gly Gly Arg Pro Ser Lys Pro Leu Gln 420 425 430 Glu Arg Leu Tyr Ser Phe Glu Phe Pro Glu Ser Pro Gly Ala Leu Leu 435 440 445 Lys Phe Leu His Thr Leu Gly Thr His Trp Asn Ile Ser Leu Phe His 450 455 460 Tyr Arg Ser His Gly Thr Asp Tyr Gly Arg Val Leu Ala Ala Phe Glu 465 470 475 480 Leu Gly Asp His Glu Pro Asp Phe Glu Thr Arg Leu His Glu Leu Gly 485 490 495 Tyr Glu Cys His Asp Glu Ser Asn Asn Pro Ala Phe Arg Phe Phe Leu 500 505 510 Ala Gly 32 514 PRT Escherichia coli 32 Met Ala Asp Ser Gln Pro Leu Ser Gly Ala Pro Glu Gly Ala Glu Tyr 1 5 10 15 Leu Arg Ala Val Leu Arg Ala Pro Val Tyr Glu Ala Ala Gln Val Thr 20 25 30 Pro Leu Gln Lys Met Glu Lys Leu Ser Ser Arg Leu Asp Asn Val Ile 35 40 45 Leu Val Lys Arg Glu Asp Arg Gln Pro Val His Ser Phe Lys Leu Arg 50 55 60 Gly Ala Tyr Ala Met Met Ala Gly Leu Thr Glu Glu Gln Lys Ala His 65 70 75 80 Gly Val Ile Thr Ala Ser Ala Gly Asn His Ala Gln Gly Val Ala Phe 85 90 95 Ser Ser Ala Arg Leu Gly Val Lys Ala Leu Ile Val Met Pro Thr Ala 100 105 110 Thr Ala Asp Ile Lys Val Asp Ala Val Arg Gly Phe Gly Gly Glu Val 115 120 125 Leu Leu His Gly Ala Asn Phe Asp Glu Ala Lys Ala Lys Ala Ile Glu 130 135 140 Leu Ser Gln Gln Gln Gly Phe Thr Trp Val Pro Pro Phe Asp His Pro 145 150 155 160 Met Val Ile Ala Gly Gln Gly Thr Leu Ala Leu Glu Leu Leu Gln Gln 165 170 175 Asp Ala His Leu Asp Arg Val Phe Val Pro Val Gly Gly Gly Gly Leu 180 185 190 Ala Ala Gly Val Ala Val Leu Ile Lys Gln Leu Met Pro Gln Ile Lys 195 200 205 Val Ile Ala Val Glu Ala Glu Asp Ser Ala Cys Leu Lys Ala Ala Leu 210 215 220 Asp Ala Gly His Pro Val Asp Leu Pro Arg Val Gly Leu Phe Ala Glu 225 230 235 240 Gly Val Ala Val Lys Arg Ile Gly Asp Glu Thr Phe Arg Leu Cys Gln 245 250 255 Glu Tyr Leu Asp Asp Ile Ile Thr Val Asp Ser Asp Ala Ile Cys Ala 260 265 270 Ala Met Lys Asp Leu Phe Glu Asp Val Arg Ala Val Ala Glu Pro Ser 275 280 285 Gly Ala Leu Ala Leu Ala Gly Met Lys Lys Tyr Ile Ala Leu His Asn 290 295 300 Ile Arg Gly Glu Arg Leu Ala His Ile Leu Ser Gly Ala Asn Val Asn 305 310 315 320 Phe His Gly Leu Arg Tyr Val Ser Glu Arg Cys Glu Leu Gly Glu Gln 325 330 335 Arg Glu Ala Leu Leu Ala Val Thr Ile Pro Glu Glu Lys Gly Ser Phe 340 345 350 Leu Lys Phe Cys Gln Leu Leu Gly Gly Arg Ser Val Thr Glu Phe Asn 355 360 365 Tyr Arg Phe Ala Asp Ala Lys Asn Ala Cys Ile Phe Val Gly Val Arg 370 375 380 Leu Ser Arg Gly Leu Glu Glu Arg Lys Glu Ile Leu Gln Met Leu Asn 385 390 395 400 Asp Gly Gly Tyr Ser Val Val Asp Leu Ser Asp Asp Glu Met Ala Lys 405 410 415 Leu His Val Arg Tyr Met Val Gly Gly Arg Pro Ser His Pro Leu Gln 420 425 430 Glu Arg Leu Tyr Ser Phe Glu Phe Pro Glu Ser Pro Gly Ala Leu Leu 435 440 445 Arg Phe Leu Asn Thr Leu Gly Thr Tyr Trp Asn Ile Ser Leu Phe His 450 455 460 Tyr Arg Ser His Gly Thr Asp Tyr Gly Arg Val Leu Ala Ala Phe Glu 465 470 475 480 Leu Gly Asp His Glu Pro Asp Phe Glu Thr Arg Leu Asn Glu Leu Gly 485 490 495 Tyr Asp Cys His Asp Glu Thr Asn Asn Pro Ala Phe Arg Phe Phe Leu 500 505 510 Ala Gly 33 329 PRT Escherichia coli 33 Met His Ile Thr Tyr Asp Leu Pro Val Ala Ile Asp Asp Ile Ile Glu 1 5 10 15 Ala Lys Gln Arg Leu Ala Gly Arg Ile Tyr Lys Thr Gly Met Pro Arg 20 25 30 Ser Asn Tyr Phe Ser Glu Arg Cys Lys Gly Glu Ile Phe Leu Lys Phe 35 40 45 Glu Asn Met Gln Arg Thr Gly Ser Phe Lys Ile Arg Gly Ala Phe Asn 50 55 60 Lys Leu Ser Ser Leu Thr Asp Ala Glu Lys Arg Lys Gly Val Val Ala 65 70 75 80 Cys Ser Ala Gly Asn His Ala Gln Gly Val Ser Leu Ser Cys Ala Met 85 90 95 Leu Gly Ile Asp Gly Lys Val Val Met Pro Lys Gly Ala Pro Lys Ser 100 105 110 Lys Val Ala Ala Thr Cys Asp Tyr Ser Ala Glu Val Val Leu His Gly 115 120 125 Asp Asn Phe Asn Asp Thr Ile Ala Lys Val Ser Glu Ile Val Glu Met 130 135 140 Glu Gly Arg Ile Phe Ile Pro Pro Tyr Asp Asp Pro Lys Val Ile Ala 145 150 155 160 Gly Gln Gly Thr Ile Gly Leu Glu Ile Met Glu Asp Leu Tyr Asp Val 165 170 175 Asp Asn Val Ile Val Pro Ile Gly Gly Gly Gly Leu Ile Ala Gly Ile 180 185 190 Ala Val Ala Ile Lys Ser Ile Asn Pro Thr Ile Arg Val Ile Gly Val 195 200 205 Gln Ser Glu Asn Val His Gly Met Ala Ala Ser Phe His Ser Gly Glu 210 215 220 Ile Thr Thr His Arg Thr Thr Gly Thr Leu Ala Asp Gly Cys Asp Val 225 230 235 240 Ser Arg Pro Gly Asn Leu Thr Tyr Glu Ile Val Arg Glu Leu Val Asp 245 250 255 Asp Ile Val Leu Val Ser Glu Asp Glu Ile Arg Asn Ser Met Ile Ala 260 265 270 Leu Ile Gln Arg Asn Lys Val Val Thr Glu Gly Ala Gly Ala Leu Ala 275 280 285 Cys Ala Ala Leu Leu Ser Gly Lys Leu Asp Gln Tyr Ile Gln Asn Arg 290 295 300 Lys Thr Val Ser Ile Ile Ser Gly Gly Asn Ile Asp Leu Ser Arg Val 305 310 315 320 Ser Gln Ile Thr Gly Phe Val Asp Ala 325 34 12 PRT Artificial Sequence Synthetic 34 Phe Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Met 1 5 10 35 12 PRT Artificial Sequence Synthetic 35 Phe Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Met 1 5 10 36 8 PRT Artificial Sequence Synthetic 36 Leu Arg Gly Ala Tyr Asn Met Met 1 5 37 28 DNA Artificial Sequence Synthetic 37 gggaattcnn gnggngcnta naanatga 28 38 12 PRT Artificial Sequence Synthetic 38 Ala Val Gly Ala Ile Leu Gly Gly Gly Gly Val Pro 1 5 10 39 12 PRT Artificial Sequence Synthetic 39 Ala Val Ala Gly Ile Leu Gly Gly Gly Gly Val Pro 1 5 10 40 7 PRT Artificial Sequence Synthetic 40 Ile Leu Gly Gly Gly Gly Val 1 5 41 28 DNA Artificial Sequence Synthetic 41 ataagcttat nanaccnccn ccnccnac 28 42 19 PRT Arabidopsis thaliana 42 Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp His Leu Arg Tyr Leu 1 5 10 15 Met Gly Gly 43 19 PRT Escherichia coli 43 Val Asn Leu Ser Asp Asp Glu Met Ala Lys Leu His Val Arg Tyr Met 1 5 10 15 Val Gly Gly 44 19 PRT Salmonella typhimurium 44 Val Asn Leu Ser Asp Asp Glu Met Ala Lys Leu His Val Arg Tyr Met 1 5 10 15 Val Gly Gly 45 19 PRT Saccharomyces cerevisiae 45 Val Asp Ile Ser Asp Asn Glu Leu Ala Lys Ser His Gly Arg Tyr Leu 1 5 10 15 Val Gly Gly 46 19 PRT Lycopersicon esculentum 46 Leu Asn Leu Ser His Asn Glu Leu Val Val Asp His Leu Lys His Leu 1 5 10 15 Val Gly Gly 47 19 PRT Cicer arietinum 47 Leu Asn Ile Ser His Asn Glu Leu Ala Val Ile His Gly Lys His Leu 1 5 10 15 Val Gly Gly 48 15 DNA Arabidopsis thaliana 48 cacctgcgtt acttg 15 49 15 DNA Arabidopsis thaliana 49 cacctgtgtt acttg 15 50 19 PRT Arabidopsis thaliana 50 Trp Asn Ile Thr Leu Phe His Tyr Arg Gly Gln Gly Glu Thr Gly Ala 1 5 10 15 Asn Val Leu 51 19 PRT Escherichia coli 51 Trp Asn Ile Ser Leu Phe His Tyr Arg Ser His Gly Thr Asp Tyr Gly 1 5 10 15 Arg Val Leu 52 19 PRT Salmonella typhimurium 52 Trp Asn Ile Ser Leu Phe His Tyr Arg Ser His Gly Thr Asp Tyr Gly 1 5 10 15 Arg Val Leu 53 19 PRT Saccharomyces cerevisiae 53 Trp Asn Leu Thr Leu Phe His Tyr Arg Asn His Gly Ala Asp Ile Gly 1 5 10 15 Lys Val Leu 54 19 PRT Lycopersicon esculentum 54 Trp Asn Ile Thr Leu Cys Arg Tyr Arg Asn Gln Gly Asp Ile Asn Ala 1 5 10 15 Ser Leu Leu 55 19 PRT Cicer arietinum 55 Trp Asn Leu Thr Leu Tyr Arg Tyr Arg Asn Gln Gly Asp Leu Lys Ala 1 5 10 15 Thr Ile Leu 56 15 DNA Arabidopsis thaliana 56 cattaccgtg gacag 15 57 15 DNA Arabidopsis thaliana 57 cattaccatg gacag 15 58 2277 DNA Arabidopsis thaliana 58 tctagaacta gtggatcccc cgggctgcag gaattcggca cgaggacggc gcaatcctct 60 ctccgtagcc acattcaccg tccatcaaaa ccagtggtcg gattcactca cttctcctcc 120 cgttctcgga tcgcagtggc ggttctgtcc cgagatgaaa catctatgac tccaccgcct 180 ccaaagcttc ctttaccacg tcttaaggtc tctccgaatt cgttgcaata ccctgccggt 240 tacctcggtg ctgtaccaga acgtacgaac gaggctgaga acggaagcat cgcggaagct 300 atggagtatt tgacgaatat actgtccact aaggtttacg acatcgccat tgagtcacca 360 ctccaattgg ctaagaagct atctaagaga ttaggtgttc gtatgtatct taaaagagaa 420 gacttgcaac ctgtattctc gtttaagctt cgtggagctt acaatatgat ggtgaaactt 480 ccagcagatc aattggcaaa aggagttatc tgctcttcag ctggaaacca tgctcaagga 540 gttgctttat ctgctagtaa actcggctgc actgctgtga ttgttatgcc tgttacgact 600 cctgagataa agtggcaagc tgtagagaat ttgggtgcaa cggttgttct tttcggagat 660 tcgtatgatc aagcacaagc acatgctaag atacgagctg aagaagaggg tctgacgttt 720 atacctcctt ttgatcaccc tgatgttatt gctggacaag ggactgttgg gatggagatc 780 actcgtcagg ctaagggtcc attgcatgct atatttgtgc cagttggtgg tggtggttta 840 atagctggta ttgctgctta tgtgaagagg gtttctcccg aggtgaagat cattggtgta 900 gaaccagctg acgcaaatgc aatggctttg tcgctgcatc acggtgagag ggtgatattg 960 gaccaggttg ggggatttgc agatggtgta gcagttaaag aagttggtga agagactttt 1020 cgtataagca gaaatctaat ggatggtgtt gttcttgtca ctcgtgatgc tatttgtgca 1080 tcaataaagg atatgtttga ggagaaacgg aacatattgg aaccagcagg ggctcttgca 1140 ctcgctggag ctgaggcata ctgtaaatat tatggcctaa aggacgtgaa tgtcgtagcc 1200 ataaccagtg gcgctaacat gaactttgac aagctaagga ttgtgacaga actcgccaat 1260 gtcggtaggc aacaggaagc tgttcttgct actctcatgc cggaaaaacc tggaagcttt 1320 aagcaatttt gtgagctggt tggaccaatg aacataagcg agttcaaata tagatgtagc 1380 tcggaaaagg aggctgttgt actatacagt gtcggagttc acacagctgg agagctcaaa 1440 gcactacaga agagaatgga atcttctcaa ctcaaaactg tcaatctcac taccagtgac 1500 ttagtgaaag atcacctgtg ttacttgatg ggaggaagat ctactgttgg agacgaggtt 1560 ctatgccgat tcacctttcc cgagagacct ggtgctctaa tgaacttctt ggactctttc 1620 agtccacggt ggaacatcac ccttttccat taccatggac agggtgagac gggcgcgaat 1680 gtgctggtcg ggatccaagt ccccgagcaa gaaatggagg aatttaaaaa ccgagctaaa 1740 gctcttggat acgactactt cttagtaagt gatgacgact attttaagct tctgatgcac 1800 tgagtttgaa gctgtggtgg ataatccaaa tctcaggaag aagaagaacc catgagagtc 1860 ttcctcgtga tcatggttgt tcttgagatt ctttagtctg ttttctctcg ggtctgtgtc 1920 tgtcggatga gcgttttagc cactgtagtt caatgagtaa cctctatttg ctacgaactc 1980 tcattcctag atcgtgggtt accttttggt ttctccaagc aatttgaggc tagcctccaa 2040 taaaaaatag tatttctagt atttgaaaaa acgctacttt cgtggtatag agaaagataa 2100 agagagagag agagagagag agagagagag agagagagag agagagagag agagagagat 2160 gctcttgata ttgctcttga tacaactcta ttattattgc tcttaatcca taatgaaagt 2220 gctttatgaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaac tcgagggggg gcccggt 2277 59 751 PRT Arabidopsis thaliana 59 Ser Arg Thr Ser Gly Ser Pro Gly Leu Gln Glu Phe Gly Thr Arg Thr 1 5 10 15 Ala Gln Ser Ser Leu Arg Ser His Ile His Arg Pro Ser Lys Pro Val 20 25 30 Val Gly Phe Thr His Phe Ser Ser Arg Ser Arg Ile Ala Val Ala Val 35 40 45 Leu Ser Arg Asp Glu Thr Ser Met Thr Pro Pro Pro Pro Lys Leu Pro 50 55 60 Leu Pro Arg Leu Lys Val Ser Pro Asn Ser Leu Gln Tyr Pro Ala Gly 65 70 75 80 Tyr Leu Gly Ala Val Pro Glu Arg Thr Asn Glu Ala Glu Asn Gly Ser 85 90 95 Ile Ala Glu Ala Met Glu Tyr Leu Thr Asn Ile Leu Ser Thr Lys Val 100 105 110 Tyr Asp Ile Ala Ile Glu Ser Pro Leu Gln Leu Ala Lys Lys Leu Ser 115 120 125 Lys Arg Leu Gly Val Arg Met Tyr Leu Lys Arg Glu Asp Leu Gln Pro 130 135 140 Val Phe Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Met Val Lys Leu 145 150 155 160 Pro Ala Asp Gln Leu Ala Lys Gly Val Ile Cys Ser Ser Ala Gly Asn 165 170 175 His Ala Gln Gly Val Ala Leu Ser Ala Ser Lys Leu Gly Cys Thr Ala 180 185 190 Val Ile Val Met Pro Val Thr Thr Pro Glu Ile Lys Trp Gln Ala Val 195 200 205 Glu Asn Leu Gly Ala Thr Val Val Leu Phe Gly Asp Ser Tyr Asp Gln 210 215 220 Ala Gln Ala His Ala Lys Ile Arg Ala Glu Glu Glu Gly Leu Thr Phe 225 230 235 240 Ile Pro Pro Phe Asp His Pro Asp Val Ile Ala Gly Gln Gly Thr Val 245 250 255 Gly Met Glu Ile Thr Arg Gln Ala Lys Gly Pro Leu His Ala Ile Phe 260 265 270 Val Pro Val Gly Gly Gly Gly Leu Ile Ala Gly Ile Ala Ala Tyr Val 275 280 285 Lys Arg Val Ser Pro Glu Val Lys Ile Ile Gly Val Glu Pro Ala Asp 290 295 300 Ala Asn Ala Met Ala Leu Ser Leu His His Gly Glu Arg Val Ile Leu 305 310 315 320 Asp Gln Val Gly Gly Phe Ala Asp Gly Val Ala Val Lys Glu Val Gly 325 330 335 Glu Glu Thr Phe Arg Ile Ser Arg Asn Leu Met Asp Gly Val Val Leu 340 345 350 Val Thr Arg Asp Ala Ile Cys Ala Ser Ile Lys Asp Met Phe Glu Glu 355 360 365 Lys Arg Asn Ile Leu Glu Pro Ala Gly Ala Leu Ala Leu Ala Gly Ala 370 375 380 Glu Ala Tyr Cys Lys Tyr Tyr Gly Leu Lys Asp Val Asn Val Val Ala 385 390 395 400 Ile Thr Ser Gly Ala Asn Met Asn Phe Asp Lys Leu Arg Ile Val Thr 405 410 415 Glu Leu Ala Asn Val Gly Arg Gln Gln Glu Ala Val Leu Ala Thr Leu 420 425 430 Met Pro Glu Lys Pro Gly Ser Phe Lys Gln Phe Cys Glu Leu Val Gly 435 440 445 Pro Met Asn Ile Ser Glu Phe Lys Tyr Arg Cys Ser Ser Glu Lys Glu 450 455 460 Ala Val Val Leu Tyr Ser Val Gly Val His Thr Ala Gly Glu Leu Lys 465 470 475 480 Ala Leu Gln Lys Arg Met Glu Ser Ser Gln Leu Lys Thr Val Asn Leu 485 490 495 Thr Thr Ser Asp Leu Val Lys Asp His Leu Cys Tyr Leu Met Gly Gly 500 505 510 Arg Ser Thr Val Gly Asp Glu Val Leu Cys Arg Phe Thr Phe Pro Glu 515 520 525 Arg Pro Gly Ala Leu Met Asn Phe Leu Asp Ser Phe Ser Pro Arg Trp 530 535 540 Asn Ile Thr Leu Phe His Tyr His Gly Gln Gly Glu Thr Gly Ala Asn 545 550 555 560 Val Leu Val Gly Ile Gln Val Pro Glu Gln Glu Met Glu Glu Phe Lys 565 570 575 Asn Arg Ala Lys Ala Leu Gly Tyr Asp Tyr Phe Leu Val Ser Asp Asp 580 585 590 Asp Tyr Phe Lys Leu Leu Met His Val Ser Cys Gly Gly Ser Lys Ser 595 600 605 Gln Glu Glu Glu Glu Pro Met Arg Val Phe Leu Val Ile Met Val Val 610 615 620 Leu Glu Ile Leu Ser Val Phe Ser Arg Val Cys Val Cys Arg Met Ser 625 630 635 640 Val Leu Ala Thr Val Val Gln Val Thr Ser Ile Cys Tyr Glu Leu Ser 645 650 655 Phe Leu Asp Arg Gly Leu Pro Phe Gly Phe Ser Lys Gln Phe Glu Ala 660 665 670 Ser Leu Gln Lys Ile Val Phe Leu Val Phe Glu Lys Thr Leu Leu Ser 675 680 685 Trp Tyr Arg Glu Arg Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg 690 695 700 Glu Arg Glu Arg Glu Arg Glu Arg Asp Ala Leu Asp Ile Ala Leu Asp 705 710 715 720 Thr Thr Leu Leu Leu Leu Leu Leu Ile His Asn Glu Ser Ala Leu Lys 725 730 735 Lys Lys Lys Lys Lys Lys Lys Lys Lys Leu Glu Gly Gly Pro Gly 740 745 750 60 712 PRT Arabidopsis thaliana 60 Leu Glu Leu Val Asp Pro Pro Gly Cys Arg Asn Ser Ala Arg Gly Arg 1 5 10 15 Arg Asn Pro Leu Ser Val Ala Thr Phe Thr Val His Gln Asn Gln Trp 20 25 30 Ser Asp Ser Leu Thr Ser Pro Pro Val Leu Gly Ser Gln Trp Arg Phe 35 40 45 Cys Pro Glu Met Lys His Leu Leu His Arg Leu Gln Ser Phe Leu Tyr 50 55 60 His Val Leu Arg Ser Leu Arg Ile Arg Cys Asn Thr Leu Pro Val Thr 65 70 75 80 Ser Val Leu Tyr Gln Asn Val Arg Thr Arg Leu Arg Thr Glu Ala Ser 85 90 95 Arg Lys Leu Trp Ser Ile Arg Ile Tyr Cys Pro Leu Arg Phe Thr Thr 100 105 110 Ser Pro Leu Ser His His Ser Asn Trp Leu Arg Ser Tyr Leu Arg Asp 115 120 125 Val Phe Val Cys Ile Leu Lys Glu Lys Thr Cys Asn Leu Tyr Ser Arg 130 135 140 Leu Ser Phe Val Glu Leu Thr Ile Trp Asn Phe Gln Gln Ile Asn Trp 145 150 155 160 Gln Lys Glu Leu Ser Ala Leu Gln Leu Glu Thr Met Leu Lys Glu Leu 165 170 175 Leu Tyr Leu Leu Val Asn Ser Ala Ala Leu Leu Leu Leu Cys Leu Leu 180 185 190 Arg Leu Leu Arg Ser Gly Lys Leu Arg Ile Trp Val Gln Arg Leu Phe 195 200 205 Phe Ser Glu Ile Arg Met Ile Lys His Lys His Met Leu Arg Tyr Glu 210 215 220 Leu Lys Lys Arg Val Arg Leu Tyr Leu Leu Leu Ile Thr Leu Met Leu 225 230 235 240 Leu Leu Asp Lys Gly Leu Leu Gly Trp Arg Ser Leu Val Arg Leu Arg 245 250 255 Val His Cys Met Leu Tyr Leu Cys Gln Leu Val Val Val Val Leu Val 260 265 270 Leu Leu Leu Met Arg Gly Phe Leu Pro Arg Arg Ser Leu Val Asn Gln 275 280 285 Leu Thr Gln Met Gln Trp Leu Cys Arg Cys Ile Thr Val Arg Gly Tyr 290 295 300 Trp Thr Arg Leu Gly Asp Leu Gln Met Val Gln Leu Lys Lys Leu Val 305 310 315 320 Lys Arg Leu Phe Val Ala Glu Ile Trp Met Val Leu Phe Leu Ser Leu 325 330 335 Val Met Leu Phe Val His Gln Arg Ile Cys Leu Arg Arg Asn Gly Thr 340 345 350 Tyr Trp Asn Gln Gln Gly Leu Leu His Ser Leu Glu Leu Arg His Thr 355 360 365 Val Asn Ile Met Ala Arg Thr Met Ser Pro Pro Val Ala Leu Thr Thr 370 375 380 Leu Thr Ser Gly Leu Gln Asn Ser Pro Met Ser Val Gly Asn Arg Lys 385 390 395 400 Leu Phe Leu Leu Leu Ser Cys Arg Lys Asn Leu Glu Ala Leu Ser Asn 405 410 415 Phe Val Ser Trp Leu Asp Gln Thr Ala Ser Ser Asn Ile Asp Val Ala 420 425 430 Arg Lys Arg Arg Leu Leu Tyr Tyr Thr Val Ser Glu Phe Thr Gln Leu 435 440 445 Glu Ser Ser Lys His Tyr Arg Arg Glu Trp Asn Leu Leu Asn Ser Lys 450 455 460 Leu Ser Ile Ser Leu Pro Val Thr Lys Ile Thr Cys Val Thr Trp Glu 465 470 475 480 Glu Asp Leu Leu Leu Glu Thr Arg Phe Tyr Ala Asp Ser Pro Phe Pro 485 490 495 Arg Asp Leu Val Leu Thr Ser Trp Thr Leu Ser Val His Gly Gly Thr 500 505 510 Ser Pro Phe Ser Ile Thr Met Asp Arg Val Arg Arg Ala Arg Met Cys 515 520 525 Trp Ser Gly Ser Lys Ser Pro Ser Lys Lys Trp Arg Asn Leu Lys Thr 530 535 540 Glu Leu Lys Leu Leu Asp Thr Thr Thr Ser Val Met Thr Thr Ile Leu 545 550 555 560 Ser Phe Cys Thr Glu Phe Glu Ala Val Val Asp Asn Pro Asn Leu Arg 565 570 575 Lys Lys Lys Asn Pro Glu Ser Ser Ser Ser Trp Leu Phe Leu Arg Phe 580 585 590 Phe Ser Leu Phe Ser Leu Gly Ser Val Ser Val Gly Ala Phe Pro Leu 595 600 605 Phe Asn Glu Pro Leu Phe Ala Thr Asn Ser His Ser Ile Val Gly Tyr 610 615 620 Leu Leu Val Ser Pro Ser Asn Leu Arg Leu Ala Ser Asn Lys Lys Tyr 625 630 635 640 Phe Tyr Leu Lys Lys Arg Tyr Phe Arg Gly Ile Glu Lys Asp Lys Glu 645 650 655 Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu 660 665 670 Arg Glu Met Leu Leu Ile Leu Leu Leu Ile Gln Leu Tyr Tyr Tyr Cys 675 680 685 Ser Ser Ile Met Lys Val Leu Tyr Glu Lys Lys Lys Lys Lys Lys Lys 690 695 700 Lys Lys Asn Ser Arg Gly Gly Pro 705 710 61 706 PRT Arabidopsis thaliana 61 Asn Trp Ile Pro Arg Ala Ala Gly Ile Arg His Glu Asp Gly Ala Ile 1 5 10 15 Leu Ser Pro Pro His Ser Pro Ser Ile Lys Thr Ser Gly Arg Ile His 20 25 30 Ser Leu Leu Leu Pro Phe Ser Asp Arg Ser Gly Gly Ser Val Pro Arg 35 40 45 Asn Ile Tyr Asp Ser Thr Ala Ser Lys Ala Ser Phe Thr Thr Ser Gly 50 55 60 Leu Ser Glu Phe Val Ala Ile Pro Cys Arg Leu Pro Arg Cys Cys Thr 65 70 75 80 Arg Thr Tyr Glu Arg Gly Glu Arg Lys His Arg Gly Ser Tyr Gly Val 85 90 95 Phe Asp Glu Tyr Thr Val His Gly Leu Arg His Arg His Val Thr Thr 100 105 110 Pro Ile Gly Glu Ala Ile Glu Ile Arg Cys Ser Tyr Val Ser Lys Arg 115 120 125 Arg Leu Ala Thr Cys Ile Leu Val Ala Ser Trp Ser Leu Gln Tyr Asp 130 135 140 Gly Glu Thr Ser Ser Arg Ser Ile Gly Lys Arg Ser Tyr Leu Leu Phe 145 150 155 160 Ser Trp Lys Pro Cys Ser Arg Ser Cys Phe Ile Cys Thr Arg Leu His 165 170 175 Cys Cys Asp Cys Tyr Ala Cys Tyr Asp Ser Asp Lys Val Ala Ser Cys 180 185 190 Arg Glu Phe Gly Cys Asn Gly Cys Ser Phe Arg Arg Phe Val Ser Ser 195 200 205 Thr Ser Thr Cys Asp Thr Ser Arg Arg Gly Ser Asp Val Tyr Thr Ser 210 215 220 Phe Ser Pro Cys Tyr Cys Trp Thr Arg Asp Cys Trp Asp Gly Asp His 225 230 235 240 Ser Ser Gly Gly Ser Ile Ala Cys Tyr Ile Cys Ala Ser Trp Trp Trp 245 250 255 Trp Phe Asn Ser Trp Tyr Cys Cys Leu Cys Glu Glu Gly Phe Ser Arg 260 265 270 Gly Glu Asp His Trp Cys Arg Thr Ser Arg Lys Cys Asn Gly Phe Val 275 280 285 Ala Ala Ser Arg Glu Gly Asp Ile Gly Pro Gly Trp Gly Ile Cys Arg 290 295 300 Trp Cys Ser Ser Arg Ser Trp Arg Asp Phe Ser Tyr Lys Gln Lys Ser 305 310 315 320 Asn Gly Trp Cys Cys Ser Cys His Ser Cys Tyr Leu Cys Ile Asn Lys 325 330 335 Gly Tyr Val Gly Glu Thr Glu His Ile Gly Thr Ser Arg Gly Ser Cys 340 345 350 Thr Arg Trp Ser Gly Ile Leu Ile Leu Trp Pro Lys Gly Arg Glu Cys 355 360 365 Arg Ser His Asn Gln Trp Arg His Glu Leu Gln Ala Lys Asp Cys Asp 370 375 380 Arg Thr Arg Gln Cys Arg Ala Thr Gly Ser Cys Ser Cys Tyr Ser His 385 390 395 400 Ala Gly Lys Thr Trp Lys Leu Ala Ile Leu Ala Gly Trp Thr Asn Glu 405 410 415 His Lys Arg Val Gln Ile Met Leu Gly Lys Gly Gly Cys Cys Thr Ile 420 425 430 Gln Cys Arg Ser Ser His Ser Trp Arg Ala Gln Ser Thr Thr Glu Glu 435 440 445 Asn Gly Ile Phe Ser Thr Gln Asn Cys Gln Ser His Tyr Gln Leu Ser 450 455 460 Glu Arg Ser Pro Val Leu Leu Asp Gly Arg Lys Ile Tyr Cys Trp Arg 465 470 475 480 Arg Gly Ser Met Pro Ile His Leu Ser Arg Glu Thr Trp Cys Ser Asn 485 490 495 Glu Leu Leu Gly Leu Phe Gln Ser Thr Val Glu His His Pro Phe Pro 500 505 510 Leu Pro Trp Thr Gly Asp Gly Arg Glu Cys Ala Gly Arg Asp Pro Ser 515 520 525 Pro Arg Ala Arg Asn Gly Gly Ile Lys Pro Ser Ser Ser Trp Ile Arg 530 535 540 Leu Leu Leu Ser Lys Arg Leu Phe Ala Ser Asp Ala Leu Ser Leu Lys 545 550 555 560 Leu Trp Trp Ile Ile Gln Ile Ser Gly Arg Arg Arg Thr His Glu Ser 565 570 575 Leu Pro Arg Asp His Gly Cys Ser Asp Ser Leu Val Cys Phe Leu Ser 580 585 590 Gly Leu Cys Leu Ser Asp Glu Arg Phe Ser His Cys Ser Ser Met Ser 595 600 605 Asn Leu Tyr Leu Leu Arg Thr Leu Ile Pro Arg Ser Trp Val Thr Phe 610 615 620 Trp Phe Leu Gln Ala Ile Gly Pro Pro Ile Lys Asn Ser Ile Ser Ser 625 630 635 640 Ile Lys Asn Ala Thr Phe Val Val Arg Lys Ile Lys Arg Glu Arg Glu 645 650 655 Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Glu Arg Cys 660 665 670 Ser Tyr Cys Ser Tyr Asn Ser Ile Ile Ile Ala Leu Asn Pro Lys Cys 675 680 685 Phe Met Lys Lys Lys Lys Lys Lys Lys Lys Lys Lys Thr Arg Gly Gly 690 695 700 Ala Arg 705 62 2304 DNA Arabidopsis thaliana CDS (1)..(1827) 62 atg ggc gag ctc ggt acc cgg gga tcc tct aga act agt gga tcc ccc 48 Met Gly Glu Leu Gly Thr Arg Gly Ser Ser Arg Thr Ser Gly Ser Pro 1 5 10 15 ggg ctg cag gaa ttc ggc acg agg acg gcg caa tcc tct ctc cgt agc 96 Gly Leu Gln Glu Phe Gly Thr Arg Thr Ala Gln Ser Ser Leu Arg Ser 20 25 30 cac att cac cgt cca tca aaa cca gtg gtc gga ttc act cac ttc tcc 144 His Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser 35 40 45 tcc cgt tct cgg atc gca gtg gcg gtt ctg tcc cga gat gaa aca tct 192 Ser Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser 50 55 60 atg act cca ccg cct cca aag ctt cct tta cca cgt ctt aag gtc tct 240 Met Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser 65 70 75 80 ccg aat tcg ttg caa tac cct gcc ggt tac ctc ggt gct gta cca gaa 288 Pro Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu 85 90 95 cgt acg aac gag gct gag aac gga agc atc gcg gaa gct atg gag tat 336 Arg Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr 100 105 110 ttg acg aat ata ctg tcc act aag gtt tac gac atc gcc att gag tca 384 Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser 115 120 125 cca ctc caa ttg gct aag aag cta tct aag aga tta ggt gtt cgt atg 432 Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met 130 135 140 tat ctt aaa aga gaa gac ttg caa cct gta ttc tcg ttt aag ctt cgt 480 Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg 145 150 155 160 gga gct tac aat atg atg gtg aaa ctt cca gca gat caa ttg gca aaa 528 Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys 165 170 175 gga gtt atc tgc tct tca gct gga aac cat gct caa gga gtt gct tta 576 Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu 180 185 190 tct gct agt aaa ctc ggc tgc act gct gtg att gtt atg cct gtt acg 624 Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr 195 200 205 act cct gag ata aag tgg caa gct gta gag aat ttg ggt gca acg gtt 672 Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val 210 215 220 gtt ctt ttc gga gat tcg tat gat caa gca caa gca cat gct aag ata 720 Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile 225 230 235 240 cga gct gaa gaa gag ggt ctg acg ttt ata cct cct ttt gat cac cct 768 Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro 245 250 255 gat gtt att gct gga caa ggg act gtt ggg atg gag atc act cgt cag 816 Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln 260 265 270 gct aag ggt cca ttg cat gct ata ttt gtg cca gtt ggt ggt ggt ggt 864 Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly 275 280 285 tta ata gct ggt att gct gct tat gtg aag agg gtt tct ccc gag gtg 912 Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val 290 295 300 aag atc att ggt gta gaa cca gct gac gca aat gca atg gct ttg tcg 960 Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser 305 310 315 320 ctg cat cac ggt gag agg gtg ata ttg gac cag gtt ggg gga ttt gca 1008 Leu His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala 325 330 335 gat ggt gta gca gtt aaa gaa gtt ggt gaa gag act ttt cgt ata agc 1056 Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser 340 345 350 aga aat cta atg gat ggt gtt gtt ctt gtc act cgt gat gct att tgt 1104 Arg Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys 355 360 365 gca tca ata aag gat atg ttt gag gag aaa cgg aac ata ttg gaa cca 1152 Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro 370 375 380 gca ggg gct ctt gca ctc gct gga gct gag gca tac tgt aaa tat tat 1200 Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr 385 390 395 400 ggc cta aag gac gtg aat gtc gta gcc ata acc agt ggc gct aac atg 1248 Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met 405 410 415 aac ttt gac aag cta agg att gtg aca gaa ctc gcc aat gtc ggt agg 1296 Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg 420 425 430 caa cag gaa gct gtt ctt gct act ctc atg ccg gaa aaa cct gga agc 1344 Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser 435 440 445 ttt aag caa ttt tgt gag ctg gtt gga cca atg aac ata agc gag ttc 1392 Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe 450 455 460 aaa tat aga tgt agc tcg gaa aag gag gct gtt gta cta tac agt gtc 1440 Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val 465 470 475 480 gga gtt cac aca gct gga gag ctc aaa gca cta cag aag aga atg gaa 1488 Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu 485 490 495 tct tct caa ctc aaa act gtc aat ctc act acc agt gac tta gtg aaa 1536 Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys 500 505 510 gat cac ctg tgt tac ttg atg gga gga aga tct act gtt gga gac gag 1584 Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu 515 520 525 gtt cta tgc cga ttc acc ttt ccc gag aga cct ggt gct cta atg aac 1632 Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn 530 535 540 ttc ttg gac tct ttc agt cca cgg tgg aac atc acc ctt ttc cat tac 1680 Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr 545 550 555 560 cat gga cag ggt gag acg ggc gcg aat gtg ctg gtc ggg atc caa gtc 1728 His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val 565 570 575 ccc gag caa gaa atg gag gaa ttt aaa aac cga gct aaa gct ctt gga 1776 Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly 580 585 590 tac gac tac ttc tta gta agt gat gac gac tat ttt aag ctt ctg atg 1824 Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met 595 600 605 cac tgagtttgaa gctgtggtgg ataatccaaa tctcaggaag aagaagaacc 1877 His catgagagtc ttcctcgtga tcatggttgt tcttgagatt ctttagtctg ttttctctcg 1937 ggtctgtgtc tgtcggatga gcgttttagc cactgtagtt caatgagtaa cctctatttg 1997 ctacgaactc tcattcctag atcgtgggtt accttttggt ttctccaagc aatttgaggc 2057 tagcctccaa taaaaaatag tatttctagt atttgaaaaa acgctacttt cgtggtatag 2117 agaaagataa agagagagag agagagagag agagagagag agagagagag agagagagag 2177 agagagagat gctcttgata ttgctcttga tacaactcta ttattattgc tcttaatcca 2237 taatgaaagt gctttatgaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaac tcgagggggg 2297 gcccggt 2304 63 609 PRT Arabidopsis thaliana 63 Met Gly Glu Leu Gly Thr Arg Gly Ser Ser Arg Thr Ser Gly Ser Pro 1 5 10 15 Gly Leu Gln Glu Phe Gly Thr Arg Thr Ala Gln Ser Ser Leu Arg Ser 20 25 30 His Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser 35 40 45 Ser Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser 50 55 60 Met Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser 65 70 75 80 Pro Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu 85 90 95 Arg Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr 100 105 110 Leu Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser 115 120 125 Pro Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met 130 135 140 Tyr Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg 145 150 155 160 Gly Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys 165 170 175 Gly Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu 180 185 190 Ser Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr 195 200 205 Thr Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val 210 215 220 Val Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile 225 230 235 240 Arg Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro 245 250 255 Asp Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln 260 265 270 Ala Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly 275 280 285 Leu Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val 290 295 300 Lys Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser 305 310 315 320 Leu His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala 325 330 335 Asp Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser 340 345 350 Arg Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys 355 360 365 Ala Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro 370 375 380 Ala Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr 385 390 395 400 Gly Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met 405 410 415 Asn Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg 420 425 430 Gln Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser 435 440 445 Phe Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe 450 455 460 Lys Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val 465 470 475 480 Gly Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu 485 490 495 Ser Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys 500 505 510 Asp His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu 515 520 525 Val Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn 530 535 540 Phe Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr 545 550 555 560 His Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val 565 570 575 Pro Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly 580 585 590 Tyr Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met 595 600 605 His 64 592 PRT Arabidopsis thaliana 64 Met Asn Ser Val Gln Leu Pro Thr Ala Gln Ser Ser Leu Arg Ser His 1 5 10 15 Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser Ser 20 25 30 Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser Met 35 40 45 Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser Pro 50 55 60 Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu Arg 65 70 75 80 Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr Leu 85 90 95 Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser Pro 100 105 110 Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met Tyr 115 120 125 Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg Gly 130 135 140 Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys Gly 145 150 155 160 Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu Ser 165 170 175 Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr Thr 180 185 190 Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val Val 195 200 205 Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile Arg 210 215 220 Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro Asp 225 230 235 240 Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln Ala 245 250 255 Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly Leu 260 265 270 Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val Lys 275 280 285 Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser Leu 290 295 300 His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala Asp 305 310 315 320 Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser Arg 325 330 335 Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys Ala 340 345 350 Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala 355 360 365 Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly 370 375 380 Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn 385 390 395 400 Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln 405 410 415 Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe 420 425 430 Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys 435 440 445 Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly 450 455 460 Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser 465 470 475 480 Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp 485 490 495 His Leu Cys Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 500 505 510 Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe 515 520 525 Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr His 530 535 540 Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro 545 550 555 560 Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr 565 570 575 Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 580 585 590 65 555 PRT Arabidopsis thaliana 65 Ile Pro Phe Ser Phe Arg Arg Arg Asn Pro Leu Ser Val Ala Thr Phe 1 5 10 15 Thr Val His Gln Asn Gln Trp Ser Asp Ser Leu Thr Ser Pro Pro Val 20 25 30 Leu Gly Ser Gln Trp Arg Phe Cys Pro Glu Met Lys His Leu Leu His 35 40 45 Arg Leu Gln Ser Phe Leu Tyr His Val Leu Arg Ser Leu Arg Ile Arg 50 55 60 Cys Asn Thr Leu Pro Val Thr Ser Val Leu Tyr Gln Asn Val Arg Thr 65 70 75 80 Arg Leu Arg Thr Glu Ala Ser Arg Lys Leu Trp Ser Ile Arg Ile Tyr 85 90 95 Cys Pro Leu Arg Phe Thr Thr Ser Pro Leu Ser His His Ser Asn Trp 100 105 110 Leu Arg Ser Tyr Leu Arg Asp Val Phe Val Cys Ile Leu Lys Glu Lys 115 120 125 Thr Cys Asn Leu Tyr Ser Arg Leu Ser Phe Val Glu Leu Thr Ile Trp 130 135 140 Asn Phe Gln Gln Ile Asn Trp Gln Lys Glu Leu Ser Ala Leu Gln Leu 145 150 155 160 Glu Thr Met Leu Lys Glu Leu Leu Tyr Leu Leu Val Asn Ser Ala Ala 165 170 175 Leu Leu Leu Leu Cys Leu Leu Arg Leu Leu Arg Ser Gly Lys Leu Arg 180 185 190 Ile Trp Val Gln Arg Leu Phe Phe Ser Glu Ile Arg Met Ile Lys His 195 200 205 Lys His Met Leu Arg Tyr Glu Leu Lys Lys Arg Val Arg Leu Tyr Leu 210 215 220 Leu Leu Ile Thr Leu Met Leu Leu Leu Asp Lys Gly Leu Leu Gly Trp 225 230 235 240 Arg Ser Leu Val Arg Leu Arg Val His Cys Met Leu Tyr Leu Cys Gln 245 250 255 Leu Val Val Val Val Leu Val Leu Leu Leu Met Arg Gly Phe Leu Pro 260 265 270 Arg Arg Ser Leu Val Asn Gln Leu Thr Gln Met Gln Trp Leu Cys Arg 275 280 285 Cys Ile Thr Val Arg Gly Tyr Trp Thr Arg Leu Gly Asp Leu Gln Met 290 295 300 Val Gln Leu Lys Lys Leu Val Lys Arg Leu Phe Val Ala Glu Ile Trp 305 310 315 320 Met Val Leu Phe Leu Ser Leu Val Met Leu Phe Val His Gln Arg Ile 325 330 335 Cys Leu Arg Arg Asn Gly Thr Tyr Trp Asn Gln Gln Gly Leu Leu His 340 345 350 Ser Leu Glu Leu Arg His Thr Val Asn Ile Met Ala Arg Thr Met Ser 355 360 365 Pro Pro Val Ala Leu Thr Thr Leu Thr Ser Gly Leu Gln Asn Ser Pro 370 375 380 Met Ser Val Gly Asn Arg Lys Leu Phe Leu Leu Leu Ser Cys Arg Lys 385 390 395 400 Asn Leu Glu Ala Leu Ser Asn Phe Val Ser Trp Leu Asp Gln Thr Ala 405 410 415 Ser Ser Asn Ile Asp Val Ala Arg Lys Arg Arg Leu Leu Tyr Tyr Thr 420 425 430 Val Ser Glu Phe Thr Gln Leu Glu Ser Ser Lys His Tyr Arg Arg Glu 435 440 445 Trp Asn Leu Leu Asn Ser Lys Leu Ser Ile Ser Leu Pro Val Thr Lys 450 455 460 Ile Thr Cys Val Thr Trp Glu Glu Asp Leu Leu Leu Glu Thr Arg Phe 465 470 475 480 Tyr Ala Asp Ser Pro Phe Pro Arg Asp Leu Val Leu Thr Ser Trp Thr 485 490 495 Leu Ser Val His Gly Gly Thr Ser Pro Phe Ser Ile Thr Met Asp Arg 500 505 510 Val Arg Arg Ala Arg Met Cys Trp Ser Gly Ser Lys Ser Pro Ser Lys 515 520 525 Lys Trp Arg Asn Leu Lys Thr Glu Leu Lys Leu Leu Asp Thr Thr Thr 530 535 540 Ser Val Met Thr Thr Ile Leu Ser Phe Cys Thr 545 550 555 66 551 PRT Arabidopsis thaliana 66 Glu Phe Arg Ser Ala Ser Asp Gly Ala Ile Leu Ser Pro Pro His Ser 1 5 10 15 Pro Ser Ile Lys Thr Ser Gly Arg Ile His Ser Leu Leu Leu Pro Phe 20 25 30 Ser Asp Arg Ser Gly Gly Ser Val Pro Arg Asn Ile Tyr Asp Ser Thr 35 40 45 Ala Ser Lys Ala Ser Phe Thr Thr Ser Gly Leu Ser Glu Phe Val Ala 50 55 60 Ile Pro Cys Arg Leu Pro Arg Cys Cys Thr Arg Thr Tyr Glu Arg Gly 65 70 75 80 Glu Arg Lys His Arg Gly Ser Tyr Gly Val Phe Asp Glu Tyr Thr Val 85 90 95 His Gly Leu Arg His Arg His Val Thr Thr Pro Ile Gly Glu Ala Ile 100 105 110 Glu Ile Arg Cys Ser Tyr Val Ser Lys Arg Arg Leu Ala Thr Cys Ile 115 120 125 Leu Val Ala Ser Trp Ser Leu Gln Tyr Asp Gly Glu Thr Ser Ser Arg 130 135 140 Ser Ile Gly Lys Arg Ser Tyr Leu Leu Phe Ser Trp Lys Pro Cys Ser 145 150 155 160 Arg Ser Cys Phe Ile Cys Thr Arg Leu His Cys Cys Asp Cys Tyr Ala 165 170 175 Cys Tyr Asp Ser Asp Lys Val Ala Ser Cys Arg Glu Phe Gly Cys Asn 180 185 190 Gly Cys Ser Phe Arg Arg Phe Val Ser Ser Thr Ser Thr Cys Asp Thr 195 200 205 Ser Arg Arg Gly Ser Asp Val Tyr Thr Ser Phe Ser Pro Cys Tyr Cys 210 215 220 Trp Thr Arg Asp Cys Trp Asp Gly Asp His Ser Ser Gly Gly Ser Ile 225 230 235 240 Ala Cys Tyr Ile Cys Ala Ser Trp Trp Trp Trp Phe Asn Ser Trp Tyr 245 250 255 Cys Cys Leu Cys Glu Glu Gly Phe Ser Arg Gly Glu Asp His Trp Cys 260 265 270 Arg Thr Ser Arg Lys Cys Asn Gly Phe Val Ala Ala Ser Arg Glu Gly 275 280 285 Asp Ile Gly Pro Gly Trp Gly Ile Cys Arg Trp Cys Ser Ser Arg Ser 290 295 300 Trp Arg Asp Phe Ser Tyr Lys Gln Lys Ser Asn Gly Trp Cys Cys Ser 305 310 315 320 Cys His Ser Cys Tyr Leu Cys Ile Asn Lys Gly Tyr Val Gly Glu Thr 325 330 335 Glu His Ile Gly Thr Ser Arg Gly Ser Cys Thr Arg Trp Ser Gly Ile 340 345 350 Leu Ile Leu Trp Pro Lys Gly Arg Glu Cys Arg Ser His Asn Gln Trp 355 360 365 Arg His Glu Leu Gln Ala Lys Asp Cys Asp Arg Thr Arg Gln Cys Arg 370 375 380 Ala Thr Gly Ser Cys Ser Cys Tyr Ser His Ala Gly Lys Thr Trp Lys 385 390 395 400 Leu Ala Ile Leu Ala Gly Trp Thr Asn Glu His Lys Arg Val Gln Ile 405 410 415 Met Leu Gly Lys Gly Gly Cys Cys Thr Ile Gln Cys Arg Ser Ser His 420 425 430 Ser Trp Arg Ala Gln Ser Thr Thr Glu Glu Asn Gly Ile Phe Ser Thr 435 440 445 Gln Asn Cys Gln Ser His Tyr Gln Leu Ser Glu Arg Ser Pro Val Leu 450 455 460 Leu Asp Gly Arg Lys Ile Tyr Cys Trp Arg Arg Gly Ser Met Pro Ile 465 470 475 480 His Leu Ser Arg Glu Thr Trp Cys Ser Asn Glu Leu Leu Gly Leu Phe 485 490 495 Gln Ser Thr Val Glu His His Pro Phe Pro Leu Pro Trp Thr Gly Asp 500 505 510 Gly Arg Glu Cys Ala Gly Arg Asp Pro Ser Pro Arg Ala Arg Asn Gly 515 520 525 Gly Ile Lys Pro Ser Ser Ser Trp Ile Arg Leu Leu Leu Ser Lys Arg 530 535 540 Leu Phe Ala Ser Asp Ala Leu 545 550 67 592 PRT Arabidopsis thaliana 67 Met Asn Ser Val Gln Leu Pro Thr Ala Gln Ser Ser Leu Arg Ser His 1 5 10 15 Ile His Arg Pro Ser Lys Pro Val Val Gly Phe Thr His Phe Ser Ser 20 25 30 Arg Ser Arg Ile Ala Val Ala Val Leu Ser Arg Asp Glu Thr Ser Met 35 40 45 Thr Pro Pro Pro Pro Lys Leu Pro Leu Pro Arg Leu Lys Val Ser Pro 50 55 60 Asn Ser Leu Gln Tyr Pro Ala Gly Tyr Leu Gly Ala Val Pro Glu Arg 65 70 75 80 Thr Asn Glu Ala Glu Asn Gly Ser Ile Ala Glu Ala Met Glu Tyr Leu 85 90 95 Thr Asn Ile Leu Ser Thr Lys Val Tyr Asp Ile Ala Ile Glu Ser Pro 100 105 110 Leu Gln Leu Ala Lys Lys Leu Ser Lys Arg Leu Gly Val Arg Met Tyr 115 120 125 Leu Lys Arg Glu Asp Leu Gln Pro Val Phe Ser Phe Lys Leu Arg Gly 130 135 140 Ala Tyr Asn Met Met Val Lys Leu Pro Ala Asp Gln Leu Ala Lys Gly 145 150 155 160 Val Ile Cys Ser Ser Ala Gly Asn His Ala Gln Gly Val Ala Leu Ser 165 170 175 Ala Ser Lys Leu Gly Cys Thr Ala Val Ile Val Met Pro Val Thr Thr 180 185 190 Pro Glu Ile Lys Trp Gln Ala Val Glu Asn Leu Gly Ala Thr Val Val 195 200 205 Leu Phe Gly Asp Ser Tyr Asp Gln Ala Gln Ala His Ala Lys Ile Arg 210 215 220 Ala Glu Glu Glu Gly Leu Thr Phe Ile Pro Pro Phe Asp His Pro Asp 225 230 235 240 Val Ile Ala Gly Gln Gly Thr Val Gly Met Glu Ile Thr Arg Gln Ala 245 250 255 Lys Gly Pro Leu His Ala Ile Phe Val Pro Val Gly Gly Gly Gly Leu 260 265 270 Ile Ala Gly Ile Ala Ala Tyr Val Lys Arg Val Ser Pro Glu Val Lys 275 280 285 Ile Ile Gly Val Glu Pro Ala Asp Ala Asn Ala Met Ala Leu Ser Leu 290 295 300 His His Gly Glu Arg Val Ile Leu Asp Gln Val Gly Gly Phe Ala Asp 305 310 315 320 Gly Val Ala Val Lys Glu Val Gly Glu Glu Thr Phe Arg Ile Ser Arg 325 330 335 Asn Leu Met Asp Gly Val Val Leu Val Thr Arg Asp Ala Ile Cys Ala 340 345 350 Ser Ile Lys Asp Met Phe Glu Glu Lys Arg Asn Ile Leu Glu Pro Ala 355 360 365 Gly Ala Leu Ala Leu Ala Gly Ala Glu Ala Tyr Cys Lys Tyr Tyr Gly 370 375 380 Leu Lys Asp Val Asn Val Val Ala Ile Thr Ser Gly Ala Asn Met Asn 385 390 395 400 Phe Asp Lys Leu Arg Ile Val Thr Glu Leu Ala Asn Val Gly Arg Gln 405 410 415 Gln Glu Ala Val Leu Ala Thr Leu Met Pro Glu Lys Pro Gly Ser Phe 420 425 430 Lys Gln Phe Cys Glu Leu Val Gly Pro Met Asn Ile Ser Glu Phe Lys 435 440 445 Tyr Arg Cys Ser Ser Glu Lys Glu Ala Val Val Leu Tyr Ser Val Gly 450 455 460 Val His Thr Ala Gly Glu Leu Lys Ala Leu Gln Lys Arg Met Glu Ser 465 470 475 480 Ser Gln Leu Lys Thr Val Asn Leu Thr Thr Ser Asp Leu Val Lys Asp 485 490 495 His Leu Arg Tyr Leu Met Gly Gly Arg Ser Thr Val Gly Asp Glu Val 500 505 510 Leu Cys Arg Phe Thr Phe Pro Glu Arg Pro Gly Ala Leu Met Asn Phe 515 520 525 Leu Asp Ser Phe Ser Pro Arg Trp Asn Ile Thr Leu Phe His Tyr Arg 530 535 540 Gly Gln Gly Glu Thr Gly Ala Asn Val Leu Val Gly Ile Gln Val Pro 545 550 555 560 Glu Gln Glu Met Glu Glu Phe Lys Asn Arg Ala Lys Ala Leu Gly Tyr 565 570 575 Asp Tyr Phe Leu Val Ser Asp Asp Asp Tyr Phe Lys Leu Leu Met His 580 585 590 68 555 PRT Arabidopsis thaliana 68 Ile Pro Phe Ser Phe Arg Arg Arg Asn Pro Leu Ser Val Ala Thr Phe 1 5 10 15 Thr Val His Gln Asn Gln Trp Ser Asp Ser Leu Thr Ser Pro Pro Val 20 25 30 Leu Gly Ser Gln Trp Arg Phe Cys Pro Glu Met Lys His Leu Leu His 35 40 45 Arg Leu Gln Ser Phe Leu Tyr His Val Leu Arg Ser Leu Arg Ile Arg 50 55 60 Cys Asn Thr Leu Pro Val Thr Ser Val Leu Tyr Gln Asn Val Arg Thr 65 70 75 80 Arg Leu Arg Thr Glu Ala Ser Arg Lys Leu Trp Ser Ile Arg Ile Tyr 85 90 95 Cys Pro Leu Arg Phe Thr Thr Ser Pro Leu Ser His His Ser Asn Trp 100 105 110 Leu Arg Ser Tyr Leu Arg Asp Val Phe Val Cys Ile Leu Lys Glu Lys 115 120 125 Thr Cys Asn Leu Tyr Ser Arg Leu Ser Phe Val Glu Leu Thr Ile Trp 130 135 140 Asn Phe Gln Gln Ile Asn Trp Gln Lys Glu Leu Ser Ala Leu Gln Leu 145 150 155 160 Glu Thr Met Leu Lys Glu Leu Leu Tyr Leu Leu Val Asn Ser Ala Ala 165 170 175 Leu Leu Leu Leu Cys Leu Leu Arg Leu Leu Arg Ser Gly Lys Leu Arg 180 185 190 Ile Trp Val Gln Arg Leu Phe Phe Ser Glu Ile Arg Met Ile Lys His 195 200 205 Lys His Met Leu Arg Tyr Glu Leu Lys Lys Arg Val Arg Leu Tyr Leu 210 215 220 Leu Leu Ile Thr Leu Met Leu Leu Leu Asp Lys Gly Leu Leu Gly Trp 225 230 235 240 Arg Ser Leu Val Arg Leu Arg Val His Cys Met Leu Tyr Leu Cys Gln 245 250 255 Leu Val Val Val Val Leu Val Leu Leu Leu Met Arg Gly Phe Leu Pro 260 265 270 Arg Arg Ser Leu Val Asn Gln Leu Thr Gln Met Gln Trp Leu Cys Arg 275 280 285 Cys Ile Thr Val Arg Gly Tyr Trp Thr Arg Leu Gly Asp Leu Gln Met 290 295 300 Val Gln Leu Lys Lys Leu Val Lys Arg Leu Phe Val Ala Glu Ile Trp 305 310 315 320 Met Val Leu Phe Leu Ser Leu Val Met Leu Phe Val His Gln Arg Ile 325 330 335 Cys Leu Arg Arg Asn Gly Thr Tyr Trp Asn Gln Gln Gly Leu Leu His 340 345 350 Ser Leu Glu Leu Arg His Thr Val Asn Ile Met Ala Arg Thr Met Ser 355 360 365 Pro Pro Val Ala Leu Thr Thr Leu Thr Ser Gly Leu Gln Asn Ser Pro 370 375 380 Met Ser Val Gly Asn Arg Lys Leu Phe Leu Leu Leu Ser Cys Arg Lys 385 390 395 400 Asn Leu Glu Ala Leu Ser Asn Phe Val Ser Trp Leu Asp Gln Thr Ala 405 410 415 Ser Ser Asn Ile Asp Val Ala Arg Lys Arg Arg Leu Leu Tyr Tyr Thr 420 425 430 Val Ser Glu Phe Thr Gln Leu Glu Ser Ser Lys His Tyr Arg Arg Glu 435 440 445 Trp Asn Leu Leu Asn Ser Lys Leu Ser Ile Ser Leu Pro Val Thr Lys 450 455 460 Ile Thr Cys Val Thr Trp Glu Glu Asp Leu Leu Leu Glu Thr Arg Phe 465 470 475 480 Tyr Ala Asp Ser Pro Phe Pro Arg Asp Leu Val Leu Thr Ser Trp Thr 485 490 495 Leu Ser Val His Gly Gly Thr Ser Pro Phe Ser Ile Thr Val Asp Arg 500 505 510 Val Arg Arg Ala Arg Met Cys Trp Ser Gly Ser Lys Ser Pro Ser Lys 515 520 525 Lys Trp Arg Asn Leu Lys Thr Glu Leu Lys Leu Leu Asp Thr Thr Thr 530 535 540 Ser Val Met Thr Thr Ile Leu Ser Phe Cys Thr 545 550 555 69 551 PRT Arabidopsis thaliana 69 Glu Phe Arg Ser Ala Ser Asp Gly Ala Ile Leu Ser Pro Pro His Ser 1 5 10 15 Pro Ser Ile Lys Thr Ser Gly Arg Ile His Ser Leu Leu Leu Pro Phe 20 25 30 Ser Asp Arg Ser Gly Gly Ser Val Pro Arg Asn Ile Tyr Asp Ser Thr 35 40 45 Ala Ser Lys Ala Ser Phe Thr Thr Ser Gly Leu Ser Glu Phe Val Ala 50 55 60 Ile Pro Cys Arg Leu Pro Arg Cys Cys Thr Arg Thr Tyr Glu Arg Gly 65 70 75 80 Glu Arg Lys His Arg Gly Ser Tyr Gly Val Phe Asp Glu Tyr Thr Val 85 90 95 His Gly Leu Arg His Arg His Val Thr Thr Pro Ile Gly Glu Ala Ile 100 105 110 Glu Ile Arg Cys Ser Tyr Val Ser Lys Arg Arg Leu Ala Thr Cys Ile 115 120 125 Leu Val Ala Ser Trp Ser Leu Gln Tyr Asp Gly Glu Thr Ser Ser Arg 130 135 140 Ser Ile Gly Lys Arg Ser Tyr Leu Leu Phe Ser Trp Lys Pro Cys Ser 145 150 155 160 Arg Ser Cys Phe Ile Cys Thr Arg Leu His Cys Cys Asp Cys Tyr Ala 165 170 175 Cys Tyr Asp Ser Asp Lys Val Ala Ser Cys Arg Glu Phe Gly Cys Asn 180 185 190 Gly Cys Ser Phe Arg Arg Phe Val Ser Ser Thr Ser Thr Cys Asp Thr 195 200 205 Ser Arg Arg Gly Ser Asp Val Tyr Thr Ser Phe Ser Pro Cys Tyr Cys 210 215 220 Trp Thr Arg Asp Cys Trp Asp Gly Asp His Ser Ser Gly Gly Ser Ile 225 230 235 240 Ala Cys Tyr Ile Cys Ala Ser Trp Trp Trp Trp Phe Asn Ser Trp Tyr 245 250 255 Cys Cys Leu Cys Glu Glu Gly Phe Ser Arg Gly Glu Asp His Trp Cys 260 265 270 Arg Thr Ser Arg Lys Cys Asn Gly Phe Val Ala Ala Ser Arg Glu Gly 275 280 285 Asp Ile Gly Pro Gly Trp Gly Ile Cys Arg Trp Cys Ser Ser Arg Ser 290 295 300 Trp Arg Asp Phe Ser Tyr Lys Gln Lys Ser Asn Gly Trp Cys Cys Ser 305 310 315 320 Cys His Ser Cys Tyr Leu Cys Ile Asn Lys Gly Tyr Val Gly Glu Thr 325 330 335 Glu His Ile Gly Thr Ser Arg Gly Ser Cys Thr Arg Trp Ser Gly Ile 340 345 350 Leu Ile Leu Trp Pro Lys Gly Arg Glu Cys Arg Ser His Asn Gln Trp 355 360 365 Arg His Glu Leu Gln Ala Lys Asp Cys Asp Arg Thr Arg Gln Cys Arg 370 375 380 Ala Thr Gly Ser Cys Ser Cys Tyr Ser His Ala Gly Lys Thr Trp Lys 385 390 395 400 Leu Ala Ile Leu Ala Gly Trp Thr Asn Glu His Lys Arg Val Gln Ile 405 410 415 Met Leu Gly Lys Gly Gly Cys Cys Thr Ile Gln Cys Arg Ser Ser His 420 425 430 Ser Trp Arg Ala Gln Ser Thr Thr Glu Glu Asn Gly Ile Phe Ser Thr 435 440 445 Gln Asn Cys Gln Ser His Tyr Gln Leu Ser Glu Arg Ser Pro Ala Leu 450 455 460 Leu Asp Gly Arg Lys Ile Tyr Cys Trp Arg Arg Gly Ser Met Pro Ile 465 470 475 480 His Leu Ser Arg Glu Thr Trp Cys Ser Asn Glu Leu Leu Gly Leu Phe 485 490 495 Gln Ser Thr Val Glu His His Pro Phe Pro Leu Pro Trp Thr Gly Asp 500 505 510 Gly Arg Glu Cys Ala Gly Arg Asp Pro Ser Pro Arg Ala Arg Asn Gly 515 520 525 Gly Ile Lys Pro Ser Ser Ser Trp Ile Arg Leu Leu Leu Ser Lys Arg 530 535 540 Leu Phe Ala Ser Asp Ala Leu 545 550 

What is claimed is:
 1. An isolated polynucleotide comprising a nucleotide sequence having substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 2. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:2.
 3. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:3.
 4. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:4.
 5. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:5.
 6. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:6.
 7. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:7.
 8. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:8.
 9. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:9.
 10. The polynucleotide in accordance with claim 1, wherein said nucleotide sequence has substantial identity to the sequence set forth in SEQ ID NO:10.
 11. A polynucleotide comprising a nucleotide sequence selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 12. A polynucleotide having a nucleotide sequence that encodes a functional, feedback-insensitive threonine dehydratase/deaminase enzyme and that hybridizes under moderately stringent conditions with a member selected from the group consisting of the nucleotide sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 13. A nucleotide sequence encoding an amino acid sequence selected from the group consisting of the amino acid sequetce set forth in SEQ ID-NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9, the sequence set forth in SEQ ID NO:10 and amino acid sequences substantially similar thereto.
 14. A method for producing cells resistant to structural analogs of isoleucine, comprising: placing into a cell a construct comprising in the 5′ to 3′ direction of transcription a promoter functional in the cell, a first nucleotide sequence that encodes a transit peptide operably attached to the promoter, a second nucleotide sequence that encodes a mutant, feedback insensitive form of threonine deaminase/dehydratase operably attached to the first sequence, and a termination region functional in the cell operably attached to the second sequence; and growing the transformed cell whereby the first and second nucleotide sequences are expressed to provide a precursor polypeptide; wherein expression of the precursor polypeptide allows the cell to be resistant to structural analogs of isoleucine.
 15. The method according to claim 14, wherein the precursor polypeptide comprises an amino acid sequence selected from the group consisting of the amino acid sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9, the sequence set forth in SEQ ID NO:10 and amino acid sequences substantially similar thereto.
 16. The method according to claim 14, wherein the cell is selected from the group consisting of a plant cell, a bacterial cell, a fungal cell and a yeast cell.
 17. A cell produced in accordance with the method of claim
 14. 18. A DNA construct comprising a promoter operably linked to a nucleotide sequence encoding a threonine dehydratase/deaminase that is substantially resistant to feedback inhibition.
 19. The DNA construct according to claim 18, wherein the nucleotide sequence has substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 20. The DNA construct according to claim 18, wherein the promoter is a plant promoter.
 21. The DNA construct according to claim 18, wherein the promoter has substantial identity to a native threonine dehydratase/deaminase promoter.
 22. A vector useful for transforming a cell, said vector comprising a nucleotide sequence having substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 23. A plant transformed with the vector of claim 22, or progeny thereof, the plant being capable of expressing the nucleotide sequence.
 24. The plant according to claim 23, the plant being selected from the group consisting of gymnosperms, rice, wheat, barley, rye, corn, potato, carrot, sweet potato, bean, pea, chicory, lettuce, cabbage, cauliflower, broccoli, turnip, radish, spinach, asparagus, onion, garlic, eggplant, pepper, celery, squash, pumpkin, zucchini, cucumber, apple, pear, quince, melon, plum, cherry, peach, nectarine, apricot, strawberry, grape, raspberry, blackberry, pineapple, avocado, papaya, mango, banana, soybean, tobacco, tomato, sorghum and sugarcane.
 25. A microorganism transformed with the vector of claim 22, or progeny thereof, the microorganism being capable of expressing the nucleotide sequence.
 26. The microorganism of claim 25, wherein said microorganism is a yeast cell.
 27. The microorganism of claim 25, wherein said microorganism is a bacterial cell.
 28. The microorganism of claim 25, wherein said microorganism is a fungal cell.
 29. A cell having incorporated therein a foreign nucleotide sequence comprising a promoter operably linked to a nucleotide sequence having substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 30. The cell according to claim 29, wherein the cell is a microorganism.
 31. The cell according to claim 29, wherein the cell is a bacterial cell.
 32. The cell according to claim 29, wherein the cell is a fungal cell.
 33. The cell according to claim 29, wherein the cell is a yeast cell.
 34. The cell according to claim 29, wherein the cell is a plant cell.
 35. A plant having incorporated into its genome a foreign DNA construct comprising a promoter operably linked to a nucleotide sequence having substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 36. A cell having incorporated into its genome a foreign nucleotide sequence encoding a threonine dehydratase/deaminase that is substantially resistant to feedback inhibition.
 37. A method comprising: incorporating into a plant's genome a DNA construct to provide a transformed plant, the construct comprising a promoter operably linked to a nucleotide sequence having substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10; wherein the transformed plant is capable of expressing the nucleotide sequence.
 38. A method comprising: providing a vector comprising a promoter operably linked to a nucleotide sequence encoding a threonine dehydratase/deaminase that is resistant to feedback inhibition, wherein the promoter regulates expression of the nucleotide sequence in a host plant cell; and transforming a target plant with the vector to provide a transformed plant, the transformed plant being capable of expressing the nucleotide sequence.
 39. The method according to claim 38, wherein the threonine dehydratase/deaminase comprises an amino acid sequence having substantial similarity to a member selected from the group consisting of the sequence set forth in SEQ ID NO: 2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 40. The method according to claim 38, wherein the nucleotide sequence has substantial identity to the nucleotide sequence of SEQ ID NO:2.
 41. A transgenic plant obtained according to the method of claim 38 or progeny thereof.
 42. A method for screening potential transformants, comprising: providing a plurality of cells, wherein at least one of the cells has in its genome an expressible foreign nucleotide sequence having substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10; and contacting the plurality of cells with a substrate comprising a toxic isoleucine structural analog; wherein cells comprising the expressible foreign nucleotide sequence are capable of growing in the substrate, and wherein cells not comprising the expressible foreign nucleotide sequence are incapable of growing in the substrate.
 43. A method for reliably incorporating a first, expressible, foreign nucleotide sequence into a target cell, comprising: providing a vector comprising a promoter operably linked to a first primary polynucleotide and a second polynucleotide comprising a nucleotide sequence having substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO: 2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10; transforming the target cell with the vector to provide a transformed cell; and contacting the cell with a substrate comprising L-O-methylthreonine; wherein successfully transformed cells are capable of growing in the substrate, and wherein unsuccessfully transformed cells are incapable of growing in the substrate.
 44. A method according to claim 43, wherein the cell is selected from the group comprising a plant cell, a yeast cell, a bacterial cell and a fungal cell.
 45. A method for growing a plurality of plants in the absence of undesirable plants, comprising: providing a plurality of plants, each having in its genome a foreign nucleotide sequence comprising a promoter operably linked to a nucleotide sequence encoding a threonine dehydratase/deaminase that is resistant to feedback inhibition; growing the plurality of plants in a substrate; and introducing a preselected amount of an isoleucine structural analog into the substrate.
 46. A method according to claim 45, wherein the nucleotide sequence has substantial identity to a member selected from the group consisting of the sequence set forth in SEQ ID NO:2, the sequence set forth in SEQ ID NO:3, the sequence set forth in SEQ ID NO:4, the sequence set forth in SEQ ID NO:5, the sequence set forth in SEQ ID NO:6, the sequence set forth in SEQ ID NO:7, the sequence set forth in SEQ ID NO:8, the sequence set forth in SEQ ID NO:9 and the sequence set forth in SEQ ID NO:10.
 47. The method in accordance with claim 45, wherein the analog is L-O-methylthreonine.
 48. A method comprising: providing a nucleotide sequence having substantial identity to the nucleotide sequence set forth in SEQ ID NO:1 or a portion thereof; and mutating the sequence so that the sequence encodes a feedback insensitive threonine dehydratase/deaminase; wherein said mutating comprises site-directed mutagenesis.
 49. The method according to claim 48, wherein the feedback insensitive threonine dehydratase/deaminase comprises an amino acid other than the wild-type at the amino acid location corresponding to location 452 of SEQ ID NO:2, and at the amino acid location corresponding to location 497 of SEQ ID NO:2.
 50. A method comprising: providing a vector comprising a promoter operably linked to a nucleotide sequence encoding a threonine dehydratase/deaminase that is resistant to feedback inhibition, wherein the promoter regulates expression of the nucleotide sequence in a host cell; and transforming a target cell with the vector to provide a transformed cell, the transformed cell being capable of expressing the nucleotide sequence. 